Abstracts & Full Text Book

Organized By

INTERNATIONAL BUFFALO FEDERATION Monterotondo, Roma, Italy

Supported By

www.wbc2019.org Abstracts & Full Text Book

Organized By

INTERNATIONAL BUFFALO FEDERATION Monterotondo, Roma, Italy 12th World Buffalo Congress Book of Abstract

Publisher : Namık Kemal University, Faculty of Agriculture Edited by : Prof. Dr. M. İhsan SOYSAL, IBF President, Organizing Committee Chair Printing Layout : Onur Nair ARDA, Tolga KOÇ Book Design : Arda ENGİN Composition : Arber Professional Congress Services

Submission and evaluation process has been handled by ARBER Professional Management System.

The organizers do not have any legal liability for the contents of the abstracts.

Indexed / Abstracted by CABI abstracts Dear delegates,

It is great honour and pleasure to welcome you to the 12th World Buffalo Congress (WBC 2019) being held 18-20 September 2019 at Istanbul. WBC 2019 has collaborated with General Directorate of Agricultural research and policy of Ministry , Agriculture and Forest of Republic of Turkey, Central Union of Water Buffalo breeder association Five significant government agencies and non-governmental organization especially and also Central union of water buffalo breeder association of Turkey as co-host of this congresses. More over these events could not be successful without support from our valuable provincial Project leader of community based water buffalo improvement program and provincial water buffalo association of Turkey . We have more than 320 participants including scientist, buffalo practitioners, breeders, and development personnel’s, graduate and under graduate students from 35 countries attend the WBC 2019 .

Organized By World buffalo population is estimated as 200 million heads present in 129 countries with over 95% of the total population in Asia producing the majority of buffalo milk and meat. Buffalo population is increased 12.5% in Asia durıng last ten years According to FAO forecast, by the year 2050 the world meat production is expected to reach twofold of present level especially in developing countries as 470 million tonnes. Swamp buffalo is known for relatively lesser milk production of around 600 kg per year and kept or draught purpose, meat production mainly. On the other hand, river buffalo is popular for it’s higher milk production around 2000 kg per lactation in improved breed of buffalo and better meat production potential. Over time buffalo rearing has shifted from the backyard to commercial farms and large business enterprises. The immense popularity of buffalo milk and meat Supported By products has ensured that buffalo production has followed in the path of the dairy cattle industry. However for this species to perform optimally under the pressure of intensive production systems, buffalo breeds have to be improved, with clear focus on the desired output. This has not yet happened enough. Buffalo, although potentially excellent for both milk and meat production, still languish in obscure conditions of poor nutrition, breeding, management and welfare. Buffalo farming is carried out in all the continents and region of world suggesting the successful adaptation of animal to the variable climate, topography as well as socio-economic conditions. Differences in climate, local geography, cropping systems and the size of farms determine the productions systems used for buffalo farming. In Italy, Mediterranean buffalo are kept on large commercial farms under modern intensive systems, primarily for producing milk, whereas river buffaloes in southwest Asian countries farms under semi intensive or extensive productıon system including swamp buffalo breeding countries mostly Southeast Asian countries. Buffalo is contributing approximately 13% to the total global milk production with India and Pakistan sharing 90% to the total global buffalo milk production. Efforts to improve the genetic potentials of the animals elsewhere are mostly government or semigovernment initiated commercial breeders associations. Among the smallholder system, which represents more than 95 % of the water buffalo population, the genetic improvement of the animals is considerably slow despite the presence of outstanding animals in the population. Dairy production system shifts more to commercial scale. Genetic improvement programme for buffalo for increasing milk production (2286 kg; 270 day) realized by establishing a national system of pedigree registration and trait measurement and recording together with reproductive technologies such as artificial insemination as shown in Italy. The many factors that constrain commercial buffalo milk production include animals’ late age at first calving, the seasonality of oestrus, and the long calving interval and dry period. Many countries had started such genetic improvement programs. Accelerated genetic improvement would be assisted through the application of molecular or genome assisted breeding. These trends needs share of experience a view by congress participants who’s applied such genetic improvement programs. The program consists of scientific and technical programs as well as social and cultural activities. The scientific and technical Organization Office programs consist of pre-congress workshop, plenary sessions" from World renowned invited speakers, 63 oral E-mail : [email protected] presentation and 85 poster presentation. First of all I would like thanks to everyone made possible 12 th World Phn : +90 312 441 0700 buffalo Congress.. It is privilege and honour for me to grace the congresses that emphasize the importance of www.arber.com.tr the buffalo production The WBC2019 provides platform to bring together not only academia but also practitioners in Buffalo world. With this platform congresses will embark on a whole process of making new discoveries and the utilizing them in to products and service for market place which wıll only made possible by this community. The theme of WBC 2019 is “efficient production”. We believe that Global warming, environmental and ecological degradation, depletion of natural resources, natural and man-made hazards, and

economic crises affects to our modes of production, and consumption, demanding for new strategies in production systems and organizations and conduct under the theme of efficient production . The congress are not only avail all participants to enrich in high standard merits, but also to open up scenario for all to enjoy the social and cultural environments before during and after the congress. You are all welcome to take this opportunity to make new collaboration for future scientific contributions.

The International Buffalo Federation (IBF) was created at the First World Buffalo Congress, that took place on, 1985, in Egypt. So far 11 World Buffalo Congress was held for each 3-4 year’s period.From 2th to 11th World Buffalo Congress was held respectively in India(1988), Bulgaria(1991), Brasil(1994), İtaly(1997), Venezuela(2001), Philipines(2005), Italy(2007), Argentina(2010), Tailand(2013), Colombia(2016) and now on 18-20 September 2019 in İstanbul ,Turkey.

In 1992 the Asian Buffalo Association (ABA), under the İnternational buffalo federation (IBF) umbrella, was established . There is an agreement between IBF and ABA to organize the respective congresses in different years and avoiding the coincidence with the World Buffalo Congress. So far 10 Asian Buffalo congress had been organized. From 1st to tenth Asian Buffalo Congress (ABC), organized by ABA, were held in, Thailand(1994); Philippines(1996) Sri Lanka, ( 2000), India, (2003).. From 1994 to date of 2013 respectively as under themes of “ Long-Term Genetic Improvement of the Buffalo (Thailand)”, Implication of changing economies in Asia on Buffalo Production (Philippines), Changing Role of the Buffalo in the New Millennium in Asia (Sri Lanka), “Role of Buffalo for Food Security and Rural Employment (India), “Economic Contribution of Buffalo in Rural Areas (China), “Buffalo Prospective Animal for Milk and Meat Enterprise (Pakistan), “Green production against global warming (Thailand) “sustainable production in the global economic World(Turkey)” . “Climate resilient buffalo production for sustainable livelihood”(India).

As alvays by any occation of congress and also in this congress we participating targeted and expected to provide a bridge among the participant from all ower the world addresses key issues relevant to buffalo production, the research and science communities, national and international regulatory bodies, policy makers and consumer organizations. The location has been chosen to reflect the role of Istanbul as a meeting point for East , West South and North as the crossroads of different continents and cultures. In addition to the main congress, there will be the PreCongress Workshop on “Reproductive Biotechnology in Buffalo” during 16th - 17th september 2019 in Veterinary Faculty of İstanbul University Cerrahpaşa located Avcılar districht of İstanbul On 21th september , You are all welcome Technical tour to Saray water buffalo farm located at Sinekli village of Silivri district of İstanbul and Ercan Taşdelen’s water buffalo farm located Gümüşpınar Village of Çatalça district of İstanbul as an example of large and small scale farm respectively from the giving impression about farming system practised point of view. This congress could not be successful without concerted efforts contributed by all staffs and organizations from the government, private sector, breeder associations all participants and with the continuous hard work of academic committee members especially our peer reviewers. I strongly believe that next days of deliberation, interaction and discussion on the different aspects of buffalo rearing during the congres will bring concrete suggestions and recommendation related to policy formulation and broadly cover the pertinent areas for future research on buffalo production and management, breeding, health and other pertinent and allied issues. I hope the congress will be a valuable and stimulating experience to you all.

I also hope that you will get a chance to enjoy the natural beauty and historical places of İstanbul where the congress is being held.I sincerely acknowledge the contribution of the co-organizers and supporters. Finally,I would like to take this opportunity to thank and congratulate the organizing secretary, subcommittee chairs, members, colleagues and faculties who have put in tireless effort into preparation for this congress, to make it a grand success. This would not have been possible without their hard work. Once again I extend my warm welcome to all of you. Thank You

Prof. Dr. M. Ihsan SOYSAL President of World Buffalo Congress Dear delegates,

World buffalo population is estimated as 200 million heads present in 129 countries with over 95% of the total population in Asia producing the majority of buffalo milk and meat. Buffalo population is increased 12.5% in Asia durıng last ten years According to FAO forecast, by the year 2050 the world meat production is expected to reach twofold of present level especially in developing countries as 470 million tonnes. Swamp buffalo is known for relatively lesser milk production of around 600 kg per year and kept or draught purpose, meat production mainly. On the other hand, river buffalo is popular for it’s higher milk production around 2000 kg per lactation in improved breed of buffalo and better meat production potential. Over time buffalo rearing has shifted from the backyard to commercial farms and large business enterprises. The immense popularity of buffalo milk and meat products has ensured that buffalo production has followed in the path of the dairy cattle industry. However for this species to perform optimally under the pressure of intensive production systems, buffalo breeds have to be improved, with clear focus on the desired output. This has not yet happened enough. Buffalo, although potentially excellent for both milk and meat production, still languish in obscure conditions of poor nutrition, breeding, management and welfare. Buffalo farming is carried out in all the continents and region of world suggesting the successful adaptation of animal to the variable climate, topography as well as socio-economic conditions. Differences in climate, local geography, cropping systems and the size of farms determine the productions systems used for buffalo farming. In Italy, Mediterranean buffalo are kept on large commercial farms under modern intensive systems, primarily for producing milk, whereas river buffaloes in southwest Asian countries farms under semi intensive or extensive productıon system including swamp buffalo breeding countries mostly Southeast Asian countries. Buffalo is contributing approximately 13% to the total global milk production with India and Pakistan sharing 90% to the total global buffalo milk production. Efforts to improve the genetic potentials of the animals elsewhere are mostly government or semigovernment initiated commercial breeders associations. Among the smallholder system, which represents more than 95 % of the water buffalo population, the genetic improvement of the animals is considerably slow despite the presence of outstanding animals in the population. Dairy production system shifts more to commercial scale. Genetic improvement programme for buffalo for increasing milk production (2286 kg; 270 day) realized by establishing a national system of pedigree registration and trait measurement and recording together with reproductive technologies such as artificial insemination as shown in Italy. The many factors that constrain commercial buffalo milk production include animals’ late age at first calving, the seasonality of oestrus, and the long calving interval and dry period. Many countries had started such genetic improvement programs. Accelerated genetic improvement would be assisted through the application of molecular or genome assisted breeding. These trends needs share of experience a view by congress participants who’s applied such genetic improvement programs. The program consists of scientific and technical programs as well as social and cultural activities. The scientific and technical programs consist of pre-congress workshop, plenary sessions" from World renowned invited speakers, 63 oral presentation and 85 poster presentation. First of all I would like thanks to everyone made possible 12 th World buffalo Congress.. It is privilege and honour for me to grace the congresses that emphasize the importance of the buffalo production The WBC2019 provides platform to bring together not only academia but also practitioners in Buffalo world. With this platform congresses will embark on a whole process of making new discoveries and the utilizing them in to products and service for market place which wıll only made possible by this community. The theme of WBC 2019 is “efficient production”. We believe that Global warming, environmental and ecological degradation, depletion of natural resources, natural and man-made hazards, and

Prof. Dr. M. Ihsan SOYSAL President of World Buffalo Congress Dear delegates,

Committees economic crises affects to our modes of production, and consumption, demanding for new strategies in Honorary Board production systems and organizations and conduct under the theme of efficient production . The congress are Antonio Borghese, not only avail all participants to enrich in high standard merits, but also to open up scenario for all to enjoy the General Secretary of International Buffalo Federation,Italy social and cultural environments before during and after the congress. You are all welcome to take this Claudia Patricia Roldan, opportunity to make new collaboration for future scientific contributions. President of 11th World Buffalo Congress,Colombia Talat Naseer Pasha, President of 6th Asian Buffalo Congress, Pakistan The International Buffalo Federation (IBF) was created at the First World Buffalo Congress, that took place on, Inderjeet Sing, 1985, in Egypt. So far 11 World Buffalo Congress was held for each 3-4 year’s period.From 2th to 11th World President of 9th Asian Buffalo Congress, India Buffalo Congress was held respectively in India(1988), Bulgaria(1991), Brasil(1994), İtaly(1997), Rangsun Parnpai, Venezuela(2001), Philipines(2005), Italy(2007), Argentina(2010), Tailand(2013), Colombia(2016) and now on President of 10th World Buffalo Congress, Thailand 18-20 September 2019 in İstanbul ,Turkey. Local Organizing Committee M. İhsan Soysal In 1992 the Asian Buffalo Association (ABA), under the İnternational buffalo federation (IBF) umbrella, was Prof. Dr., Namık Kemal University, Congress Chair established . There is an agreement between IBF and ABA to organize the respective congresses in different Hüseyin Yılmaz years and avoiding the coincidence with the World Buffalo Congress. So far 10 Asian Buffalo congress had President of Central Water Buffalo Breeders Association' of Turkey been organized. From 1st to tenth Asian Buffalo Congress (ABC), organized by ABA, were held in, Hadi Kayhan General Secretary Central Water Buffalo Breeders Association' of Turkey Thailand(1994); Philippines(1996) Sri Lanka, ( 2000), India, (2003).. From 1994 to date of 2013 respectively as Ahmet Yavuz Karaca under themes of “ Long-Term Genetic Improvement of the Buffalo (Thailand)”, Implication of changing İstanbul Provincial Director of Ministry of Food Agriculture and Livestock economies in Asia on Buffalo Production (Philippines), Changing Role of the Buffalo in the New Millennium in E. Kemal Gürcan Asia (Sri Lanka), “Role of Buffalo for Food Security and Rural Employment (India), “Economic Contribution of Assoc. Prof. Dr., Namık Kemal University, Congress Vice-Chair Buffalo in Rural Areas (China), “Buffalo Prospective Animal for Milk and Meat Enterprise (Pakistan), “Green Emel Özkan Assoc. Prof. Dr., Namık Kemal University, Congress Vice-Chair production against global warming (Thailand) “sustainable production in the global economic World(Turkey)” . Serdar Genç “Climate resilient buffalo production for sustainable livelihood”(India). Ass.Prof.Dr., Ahi Evran University, Congress Vice-Chair

As alvays by any occation of congress and also in this congress we participating targeted and expected to Local Scientific Committee provide a bridge among the participant from all ower the world addresses key issues relevant to buffalo Şefik Kurultay, Prof. Dr., Namık Kemal University production, the research and science communities, national and international regulatory bodies, policy makers İsmail Yılmaz, Prof. Dr., Namık Kemal University and consumer organizations. The location has been chosen to reflect the role of Istanbul as a meeting point for Ahmet İstanbulluoğlu, Prof. Dr., Dean of Faculty of Agriculture of Namık Kemal University Muhittin Özder, Prof. Dr., Namık Kemal University East , West South and North as the crossroads of different continents and cultures. In addition to the main Mehmet Demirci,Prof. Dr., Namık Kemal University congress, there will be the PreCongress Workshop on “Reproductive Biotechnology in Buffalo” during Özkan Kayacan, Ministry of Agriculture and Forestry 16th - 17th september 2019 in Veterinary Faculty of İstanbul University Cerrahpaşa located Avcılar districht of Ali Ayar, Dr., Ministry of Agriculture and Forestry İstanbul On 21th september , You are all welcome Technical tour to Saray water buffalo farm located at Sinekli Bekir Ankaralı, Dr., Ministry of Agriculture and Forestry village of Silivri district of İstanbul and Ercan Taşdelen’s water buffalo farm located Gümüşpınar Village of Serhat Pabuccuoğlu, Prof. Dr., İstanbul University-Cerrahpasa Çatalça district of İstanbul as an example of large and small scale farm respectively from the giving impression Mustafa Tekerli, Prof. Dr., Afyon Kocatepe University Özel Şekerden, Prof. Dr., Mustafa Kemal University about farming system practised point of view. This congress could not be successful without concerted efforts Serhat Alkan, Prof. Dr., İstanbul University-Cerrahpasa contributed by all staffs and organizations from the government, private sector, breeder associations all Ömür Koçak, Prof. Dr., İstanbul University-Cerrahpasa participants and with the continuous hard work of academic committee members especially our peer reviewers. Güven Kaşıkçı, Prof. Dr., İstanbul University-Cerrahpasa I strongly believe that next days of deliberation, interaction and discussion on the different aspects of buffalo Zehra Sarıçiçek, Prof. Dr., Ankara University rearing during the congres will bring concrete suggestions and recommendation related to policy formulation Sezen Arat, Prof. Dr., Namık Kemal University and broadly cover the pertinent areas for future research on buffalo production and management, breeding, Gürsel Dellal, Prof. Dr., Ankara University Cengiz Elmacı, Prof. Dr., Uludağ University health and other pertinent and allied issues. I hope the congress will be a valuable and stimulating experience Alper Yılmaz, Prof. Dr., İstanbul University to you all. İsmail Abaş, Prof. Dr., İstanbul University Ersin Şamlı, Prof. Dr., Namık Kemal University I also hope that you will get a chance to enjoy the natural beauty and historical places of İstanbul where the Alper Önenç, Prof. Dr., Namık Kemal University congress is being held.I sincerely acknowledge the contribution of the co-organizers and supporters. Finally,I Fisun Koç, Assoc. Prof. Dr., Namık Kemal University would like to take this opportunity to thank and congratulate the organizing secretary, subcommittee chairs, Eser Kemal Gürcan, Assoc. Prof. Dr., Namık Kemal University Levent Özdüven, Prof. Dr., Namık Kemal University members, colleagues and faculties who have put in tireless effort into preparation for this congress, to make it a Levent Coşkuntuna, Assoc. Prof. Dr., Namık Kemal University grand success. This would not have been possible without their hard work. İsmail Kaya, Prof. Dr., Ondokuz Mayıs University Once again I extend my warm welcome to all of you. Y. Tuncay Tuna, Asst. Prof. Dr., Namık Kemal University Thank You Hüseyin Erdem, Asst. Prof. Dr., Ondokuz Mayıs University İrfan Daşkıran, Assoc.Prof.Dr., Ministry of Agriculture and Forestry Prof. Dr. M. Ihsan SOYSAL Mustafa Küçükkebapçı, Dr., Bandırma Livestock Research Institute Özden Sarıkaya, Ministry of Agriculture and Forestry President of World Buffalo Congress Yusuf Kaplan, Ministry of Agriculture and Forestry Sezer Oz, Dr., Ministry of Agriculture and Forestry Kürşat Alkoyak, Dr., Ministry of Agriculture and Forestry KEYNOTE SPEAKERS Sustainable Models Based on Energy Produced by Animal Draft

Antonio BORGHESE1, Antonio PERRONE2

1International Buffalo Federation, Italy 2Masseria Coppola Farm, Italy [email protected], [email protected]

- The theme of the 12th World Buffalo Congress is “Efficient Production for the World”. Therefore the first question is: What is the system for the best efficient production in buffalo livestock? What will be the future?

- We find in the world two ways: 1, intensive models with mechanization, as in Italy and in Europe, where all the activities are mechanized, as production of the feeding stuff, animal feeding, manure treatment, buffalo milking parlour until the use of milking robot; this process is moved by the must to reduce the cost of the workers; 2, extensive models where the buffalo is used as priority for draught power, and after that, to give also milk, meat, manure for fuel, horns, leather, bones. We can look at countries as China, Thailand, Bangladesh, Vietnam, where buffalo is used as draft animal in rice fields, or Iraq, Sumatra, Amazonia, where buffalo lives in marshlands, because only buffalo can swing and survive in the swamp areas, feeding canes, reed, and other water forages.

- Now the first problem for the survive of the Planet is the climatic change, where the greatest responsibility is due to the CO2 emission by the use of fossil energy, particularly oil, to move cars and to produce electricity, and by intensive livestock too. Therefore European Union is encouraging research projects to reduce pollution and CO2 emission in the livestock. Many farmers are coming back, also in Italy, to animal draft for cultivating lands, particularly in marginal areas, and for other agrarian activities. What have we to suggest to developing countries?

- At the moment about 250 million working animals provide the draught power to approximately 28% of the world arable land, equivalent to the 52% of the total cropping land in developing countries. The draught animal power is the only source of renewable energy totally self produced in the rural villages. Then its use does not require the supply of foreign currency, nor energy produced by fossil, responsible of climatic change.

- We find buffaloes used in the world not only for ploughing the agricultural lands, but also running other devices, such mechanical pumps, lifting water and seed crushers. In the remote villages, far from electric grid and from an economy supply of fuel, the use of draught animals appears feasible, economic and reliable. The draught animal power represents an intermediate technology from manual labor toward the introduction of the mechanization. The use of draught animal power can perfectly substitute the work of women and children now very often engaged in the villages, in the water lifting and seed crushing.

- An electricity economically generated by animal draft can be used for the above mentioned purposes, to extend the light hours for student education and for other economic and social activities. The animal welfare is not compromised by this kind of work. Millennia of selection have conducted to strong and patient animals, that do not suffer for a monotonous work, and this is the reason for which working animals are still hundreds of million in rural areas.

- Several experiments for the generation of electricity through the utilization of animal traction have been conducted in several countries in the last decades. An updated bibliography is available in www.wedap.eu/rcb.html This bibliography shows the several works published on the scientific magazines on the matter.

- Following the specific literature and test performed by WEDAP in the Masseria Coppola, Taranto, Italy, it is possible to affirm that the draught animal powered generators seems to be competitive with analogous photovoltaic plant of 8 kwp, also in terms of cost: 0.13 versus 0.01 euro for each kilowatt/hour. The draught power is more efficient at any hour, with any sun and weather conditions and as transportability and production on demand, with less cost of purchase in comparison with photovoltaic plants.

- The potential of these technologies is enormous if one thinks about the number of the working animals present in the remote villages with a prevailing rural economy. As we read in the official FAO texts, it is still a matter of hundreds of millions of active animals in agriculture. This important issue requests the building of a “working buffalo network” whose focal point could be our International Buffalo Federation. The mission of the network should be the assessment of the local research resources that could match the aims of the various governmental departments engaged in the cooperation to the development.

1 KEYNOTE SPEAKERS Modern Processing Techniques for Improving Buffalo Meat Sector of Pakistan

Talat N. PASHA

University of Veterinary and Animal Science Lahore, Pakistan [email protected]

Water buffalo is equally important as cattle in many regions and playing a vital role for socio-economic development of rural economy in Asian regions. In Pakistan buffalo supplies almost 61% of total milk and almost half of total beef production. However, potential of buffalo for meat and milk production has not been fully utilised. Our research has focused on better utilisation of male buffalo calves for meat production and use of post slaughter technologies to improve buffalo meat quality to make it more acceptable in domestic and international markets. We evaluated growth rate of veal calves (14 days old) by feeding only milk, milk replacer and calf starter for a period of 30 and 40 weeks and it was found that they gained an average of 127 and 195 kg weight during respective rearing periods which produced desirable pink veal. In another project, Buffalo calves were reared to an age of 18 months and the beef was marketed at urban centres for development of sustainable buffalo calves value chain in Pakistan. In one more study, young (18 months) and old (+10 years) age buffalo meat was compared and it was found that ageing at 14 days can reduce toughness of old age meat closer to fresh young buffalo meat. There remains a public perception that buffalo meat is tougher and of inferior quality than beef. We compared 26 months old buffalo and cattle meat and it was found that there was no significant difference between WBSF values and lightness L* values of both meats. However, cattle beef had higher redness a* and chroma C* value as compared to buffalo meat but the difference was undetectable subjectively. Electrical Stimulation (ES) and Tender Stretch method (TS) was used to evaluate their effect on meat tenderness. Results indicated that ES and TS individually and in combination improved buffalo meat tenderness. Rearing buffalo calves to higher weight group can enhance total veal production and applying post-slaughter technologies like ageing and ES can improve buffalo meat quality, while, no difference has been observed in meat tenderness of cattle and buffalo meat.

2 KEYNOTE SPEAKERS Recent Breakthrough in Modern Buffalo Production Systems In India

Inderjeth SINGH1, Ashok Kumar BALHARA2

1Animal Husbandry, Punjab Livestock Complex, India 2ICAR-Central Institute for Research on Buffaloes, India [email protected], [email protected]

Buffaloes in India are spread over almost all parts of the country with varying population density. Majority (>70%) of the population is however concentrated in the north and western states where most of the milch breeds of buffaloes are found viz. in Haryana, Punjab, Uttar Pradesh, Rajasthan, Gujarat and Maharashtra. During the last 10 years, when cattle population is on the decline, there has been continuous growth of this species in this region at the rate of close to 2% per annum as against the average growth rate of approximately 1% in rest of the country. Although larger commercial farms are also coming up, it will take time for these to get established and show the progress. In such a scenario, the responsibility to improve the buffalo germplasm and husbandry has been taken-up by the respective governments, through establishing demonstration farms, bull-mother farms, semen freezing laboratories, disease testing laboratories as well as AI centres and vaccine production units. The country at present has 59 (37 ‘A’ graded, 14 ‘B’ graded, 01 ‘A’ graded and 7 non-graded) semen stations producing 160million doses of frozen semen. The viability of such enterprises in government sector is not always guaranteed and many a times they suffer from government apathy and under-performance due to political or local administrative reasons. There is also a huge focus on production of breeding bulls in public private partnership (PPP) mode using pedigree selection and progeny testing programs. Under such circumstances the reliability of breeding and production data can be increased using molecular tools like parentage verification. These tools can be used to evaluate valuable animals pre- and post-sale, to confirm identity and the offsprings genetically against all possible parents for establishing whether the calf is the result of AI or natural service. The National Dairy Development Board (NDDB) in India has taken initiative to launch SNP based chip for parentage verification in Murrah buffaloes. The future of all livestock breeding programs is shifting towards genomic selection using molecular markers and related tools. The buffalo production systems has top cover a lot of ground to come at par with the modern breeding practices beyond the artificial insemination, which has now covered large populations of the species now.

3 KEYNOTE SPEAKERS Development of Feed Resources, Rumen Microbiome, Fermentation Efficiency, Mitigation of Methane for The Sustainable Buffalo Productivity

Metha WANAPAT

Animal Science, Faculty of Agriculture, Khon Kaen University, Thailand [email protected]

Livestock production plays a crucial role, which extends beyond its traditional supply of meat and milk. Buffaloes (Bubbalus bubbalis) are used for multiple purposes as draft power, means of transportation, capital, credit, meat, milk, social value, hides, and source of organic fertilizer for seasonal croppings. Buffaloes can utilize agricultural crop residues and farm by-product that are abundantly available. Feed resource are scarce in both quantity and quality especially during the long dry season which results in low productivity of livestock. Feed resources are major components of animal production especially among small-holder farmers. Four main categories of feeds are potentially available for use on farms: foliages (native and improved grasses, legumes and fodder trees), crop residues, agro-industrial by-products and non-conventional feed resources.

The rumen has an important role and function in preparing fermentation end-products for biosynthetic process of buffaloes. It is therefore essential that the rumen is healthy and is able to establish an optimum ecology in order to perform well with regards to rumen microbiomes (bacteria, protozoa and fungi), pH, substrate (e.g. roughage, energy, effective fibre etc.), fermentation end-products (NH3-N and VFAs), and microbial protein synthesis, VFAs, particularly propionate (C3), acetate (C2), and butyrate (C4), are major source of energy, and allow the synthesis of glucogenic and lipogenic compounds, while NH3-N is an essential source of nitrogen for microbial protein synthesis. Buffaloes depend greatly on fermentation end-products to support meat and milk production, arisen from the anaerobic fermentation (Embden-Meyerhof Parnas Pathway) by rumen microbiomes. These fermentation end-product (volatile fatty acids, VFA; ammonia nitrogen, NH3 etc.), can greatly impact on rumen ecology especially the pH. The decline of rumen pH can be attributed by the accumulation of VFA and lactic production. When rumen pH declines from 6.3 especially below 6.0-5.6, the rumen microorganisms would be consequently impacted, mostly on the cellulolytic bacteria, creating acidic environment in the rumen. Phytonutrients containing feeds or wastes, as well as plant secondary compounds have been reported to exert their effects on rumen microbiomes and fermentation by maintaining rumen pH (6.0-6.3), suppressing protozoal population, hence mitigating methane production. Currently, a number of feeds and/or fruit wastes containing phytonutrients such as condensed tannins (CT) and saponins (SP) have been used in the concentrate mixtures or as pellet supplements. Research conducted using fruit wastes such as mangosteen peel, rambutan peel, dragon fruit peel, banana flower powder, grape pomace; pellets with combinations namely BAFLOP and MARABAC have been reported to influence the rumen ecology and improve fermentation, by buffering rumen pH, increased propionic acid, microbial protein synthesis, as well as nutrient digestibilities. Nevertheless more research regarding the use of rumen enhancer containing phytonutrients from plants and fruit wastes are highly encouraged to provide more practical information. Furthermore, the food-feed-system (FFS) has been potentially implemented as it can provide year-round feed availability and utilization for ruminant production enhancement. Furthermore, strategy to combine both conventional and unconventional feed resources especially agricultural crop-residues and agro-industrial by-products are utmost important for the sustainable and efficient buffalo production, as well as to ensure the supply of meat and milk from buffalo for the global increasing population.

4 KEYNOTE SPEAKERS The Buffalo Genome and The Application of Genomics to Buffalo Breeding

John L. WILLIAMS1, Wai Yee LOW1, Daniela IAMARTINO2, Timothy P.l. SMITH3, Lynsey K. WHITACRE4, Paolo Ajmone MARSAN5, Sara ALBARELLA6, Derek M. BICKHART7, Licia COLLI8, Ezequiel L. NICOLAZZI9, James M. REECY10, Robert D. SCHNABEL11, Rick TEARLE1, Alessio VALENTINI12, Curtis P. VAN TASSELL13

1The Davies Research Centre, School of Animal and Veterinary Sciences, Australia 2AIA-LGS Associazione Italiana Allevatori, Italy 3US Meat Animal Research Center, ARS USDA, Clay Center, United States 4Biozyme Inc., 6010 Stockyards Expressway, United States 5Department of Animal Science, Food and Nutrition, Italy 6Department of Veterinary Medicine and Animal Production, Italy 7Cell Wall Biology and Utilization Laboratory, United States 8Department of Animal Science, Food and Nutrition, Università Cattolica del Sacro Cuore, Italy 9Fondazione Parco Tecnologico Padano, Italy 10Iowa State University, United States 11Division of Animal Science and Informatics Institute, United States 12University of Tuscia, Viterbo, Italy 13Animal Genomics and Improvement Laboratory, United States [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

A good reference genome is necessary to understand the biology of a species, manage genetic diversity and, in the case of livestock, to apply genomic selection for genetic improvement. Rapid innovations in sequencing technologies [1] have enabled us to create a high quality chromosome-level assembly of the water buffalo (Bubalus bubalis) [2]. Availability of whole genome sequence enables us to explore the genome structure [3,4,5], the function of genes and how they relate to phenotypes. We have demonstrated the utility of genome sequencing in an investigation of a genetic disease. Transverse hemimelia, which causes unilateral or bilateral hindlimb malformation recently appeared in Mediterranean River buffalo. To identify the genes responsible for Transverse hemimelia, we sequenced 11 affected buffaloes and compared the genome sequences with 14 normal buffaloes [6]. This analysis suggests that there are several genetic loci involved, but implicate variations in the genes SMARCA4 and WNT7A as the main drivers necessary for the manifestation of the disease.

One of the most interesting aspects of the genomic revolution for the breeding industry is the possibility to predict a “direct genomic breeding value” (DGV) using genome-wide DNA markers. To create a panel of single nucleotide polymorphic markers, suitable for use in most buffalo breeds, we sequenced the genomes of buffaloes from several breeds [7]. Millions of sequence variants were identified from which 90,000 single nucleotide polymorphisms were selected to createan Axiom® Buffalo 90K Genotyping Array. Between 60,000 and 70,000 of these SNP are polymorphic in all riverine breeds of water buffalo and 20,000 to 25,000 are polymorphic in swamp buffalo. These are sufficient for carrying out genome-wide association studies, identifying signatures of selection [8], developing genomic breeding value predictions [9] and exploring the genetic diversity of buffalo populations [10].

[1] Koren, S., et al,. (2018). Nature Biotechnology https://www.nature.com/articles/nbt.4277 [2] Low, W.Y., et al., (2019). Nature Communication 10:260. doi /10.1038/s41467-018-08260-0 [3] Liu, S., et al., (2019). Funct Integr Genomics https://doi.org/10.1007/s10142-019-00657-4 [4] Whitacre, L.K., et al. (2017). Nature Scientific Reports 7: 39719. doi: 10.1038/srep39719 [5] Iamartino, D., et al. (2017). PLoS ONE 12(10): e0185220. doi. org/10.1371/journal.pone.0185220 [6] Colli, L., et al. (2018). Front. Genet. 9:53. doi: 10.3389/fgene.2018.00053.

5 KEYNOTE SPEAKERS Keys to Improving Fertility of Anestrus Buffaloes Round the Year: An Example From Nepal

Bhuminand DEVKOTA

Department of Theriogenology, Faculty of Animal Science, Veterinary Science and Fisheries, Agriculture and Forestry University, Nepal [email protected]

Buffalo (Bubalus bubalis) production is the backbone of rural economy of many countries in the world. In Nepal, buffaloes are reared as dual-purpose livestock that contribute hugely in milk and meat production with more than two third of the total milk and half of the meat in the country coming from buffalo. However, seasonal reproductive pattern and infertility issue of anestrus characterized by silent cyclicity and acyclicity hinder their production potentiality. In order to improve the fertility of anestrus buffaloes round the year, we have focused on understanding the cyclic pattern of buffaloes during good, transition and low breeding seasons and investigated the factors affecting the pregnancy outcome while treating with novel hormones or applying the FTAI protocols during different seasons. One of our field study revealed 60% pregnancy outcome during good breeding season while giving PG treatment to buffaloes with anestrus due to silent cyclicity in which poor body condition score (BCS), sub-normal levels of some blood metabolic parameters and gastrointestinal parasitic infection were associated to minimize pregnancy. In a study when CIDR co-synch protocol for FTAI was applied in anestrus buffaloes, buffaloes having poor BCS during low season failed to conceive, however, 42.9% pregnancy during this season and 66.7% pregnancy during good season was obtained in good BCS buffaloes. Recently, we applied ovsynch protocol during good season and CIDR co-synch protocol during transition and low seasons as silent ovulation during good season and acyclicity during other two seasons are the major causes of anestrus. Applying these protocols, one study investigated the effect of blood metabolic parameters and stress factor, cortisol. It was found that ovsynch treatment during good season produced 42.9% pregnancy whereas CIDR co-synch protocol produced 50.0% pregnancy during transition and 28.6% during poor season. This study revealed blood cholesterol and cortisol levels were different (p<0.05) among three seasons and poor BCS was associated to pregnancy failure. Lately, we developed the holistic approach of improving management and nutrition along with lowering the environmental stress prior to applying those FTAI protocols in anestrus buffaloes that improved the pregnancy, particularly during low season. Thus, it was understood that novel FTAI protocols can be effectively applied to improve fertility of anestrus buffaloes round the year given the nutritional, management and stress factors are addressed.

6 ORAL ABSTRACTS A Risk Assessment Study of Poisoning by Bacillus Cereus in Buffalo Mozzarella Cheese

Angela Michela Immacolata MONTONE, Andrea MANCUSI, Federico CAPUANO, Orlandina DI MARO, Daniela CRISTIANO experimental zooprophylactic institute of the south, Italy [email protected], [email protected], [email protected], [email protected], [email protected]

Bacillus cereus group (B.c.) comprises several spoilage bacteria, some of them responsible of food borne disease and infections. Two food-borne illness are caused by Bacillus cereus in humans: a diarrheal disease, associated with cytotoxin K, enterotoxin FM, hemolysin BL and a non hemolytic enterotoxin, and an emetic syndrome, associated with cereulide toxin. Due to the heat resistance of Bacillus cereus and of its capability to grow in milk, Bacillus cereus should be considered as potentially hazardous in dairies products. The risk assessment of Bacillus cereus poisoning for the consumption of buffalo mozzarella cheese was performed in this study. Three hundred and forty mozzarella samples were analyzed for Bacillus cereus. enumerations. Bacillus cereus isolates were characterized for the presence of hblA, hblC, hblD, nheA, nheB, nheC, cytK, entFM and ces genes, related to the production of toxins. Eighty-six out of the 340 samples (25.3%) were found contaminated by Bacillus cereus strains, with values ranging from 2.2x102 to 1.6x106 CFU/g. Ces gene characterized only one of the 89 strains, while the other virulence factors analyzed were highlighted in 88 (nhe A,B,C), 87 (entFM), 41 (cytk) and 33 (hblA,C,D) of the Bacillus cereus isolates, respectively. Overall, eight different molecular profiles were identified, five of them with virulence characteristics.

The analysis of the results highlight a risk of poisoning by Bacillus cereus overall low. Indeed, only 6 (1.8%) out the 340 samples analyzed showed a level of contamination higher than 105 Bacillus cereus CFU/g. The combined evaluation of both, Bacillus cereus concentration and virulotype, highlight that only three (0.9%) mozzarella samples showed simultaneously high bacterial load (>106 CFU/g) and Bacillus cereu. isolated with hazardous virulotype; while other five samples (1.5%) harbored over 104 hazardous Bacillus cereus/g isolates.

We thank all the operating units involved and our administration that have allowed this work

[1] Beecher, D.J. & Wong, A.C. 1997. Tripartite hemolysin BL from Bacillus cereus. Hemolytic analysis of component interactions and a model for its characteristic paradoxical zone phenomenon. J. Biol. Chem., 272(1), 233-239. [2] EFSA & ECDC. 2013. The European Union Summary Report on Trends and Sources of Zoonoses, Zoonotic Agents and Food-borne Outbreaks in 2011. [3] Martinelli, D., Fortunato, F., Tafuri, S., Cozza, V., Chironna, M., Germinario, C., Pedalino, B. & Prato, R. 2013. Lessons learnt from a birthday party: a Bacillus cereus outbreak, Bari, Italy, January 2012. Ann. Ist. Sup. Sanità 49(4), 391-394.

7 ORAL ABSTRACTS Different Types of Milk Flow Profiles and Economic Impact

Carlo BOSELLI1, Antonio BORGHESE2

1Istituto Zooprofilattico Sperimentale del Lazio e della Toscana M. Aleandri, Italy 2International Buffalo Federation, Italy [email protected], [email protected]

A general definition of milkability is the ability of a female to give a regular, complete and rapid milk secretion by the mammary gland in response to a proper milking technique. Milk yield and milkability can be observed trough the milk flow profiles, both are important parameters to be recorded and evaluated, in fact they give various advices on milking management. In order to evaluate individual responses at the mechanical milking, 2,182 milk flow curves were recorded with electronic milkmeter (Lactocorder®), from 2,182 Mediterranean Italian buffalo cows in different parity and days of lactation. Statistical analysis has allowed to classify 9 different milk emission patterns. Average milk yield was 3.79±0.04 kg/head/milking, while maximum flow and average flow were 1.22±0.01 kg/min and 0.78±0.01 kg/min respectively. Average total milking machine time was higher 10.82±0.08 min, respect the time of milk ejection: the principal milking time was very low with only 4.05±0.05 min. The most represented milk flow curves were the type 3 (25.94%) with a ratio between plateau and decreasing phase near to one (1.32 vs 1.66 min), followed by type 6 (17.78%) with a very long plateau phase, while the less represented were type 1 (4.35%) characterized by long milk letdown time and incline time and low milk flow. Type 9, unclassified, was present with 5.04 % with very poor milk yield (1.14 kg).

We found 53 bimodal curves in type 4, the most high, that are representative of very short teat canals and of very high milk flow. Therefore, as type 6 is more productive in terms of milk yield, type 4 is more economic in terms of milk time. We underline that these data are valid for Mediterranean Italian breed, could be interesting to compare them with other dairy buffalo breeds; after that in Italy we can select the dairy buffaloes according curves of type 5, 6 and 4, to obtain milking buffaloes with the best milk yield and reduced milk time for the most favorable emission patterns, with positive economic impact on the farm, particularly increasing milk production and reducing milking cost.

[1] Boselli C., Rosati R., Giangolini G., Arcuri S., Fagiolo A., Ballico S., Borghese A., 2004. Milk flow measurements in buffalo herds. Proc. of the Seventh World Buffalo Congress, Manila, Philippines, 20-23 Oct.: 244-246. [2] C. Boselli, M.M. Mazzi, A. Borghese, G.M. Terzano, G. Giangolini, F. Filippetti, S. Amatiste, R. Rosati: Milk flow curve and teat anatomy in mediterranean Italian buffalo cows. Revista Veterinaria 01/2010; 21:573-578. [3] Antonio Borghese, Carlo Boselli, Remo Rosati: Lactation Curve and Milk Flow. Buffalo Bulletin 01/2013; 32:334-350. [4] Maria Caria, Carlo Boselli, Lelia Murgia, Remo Rosati, Antonio Pazzona: Effect of vacuum level on milk flow traits in Mediterranean Italian buffalo cow. 01/2012; 11(1)., DOI:10.4081/ijas.2012.e25 [5] S. Ambord, C.S. Thomas, A. Borghese, M. Mazzi, C. Boselli, R.M. Bruckmaier: Teat anatomy, vacuum to open the teat canal, and fractionized milk composition in Italian buffaloes. Milchwissenschaft 01/2009; 64(4):351-353. A.Borghese, M. Mazzi, R. Rosati, C. Boselli: Milk flow pictures in Mediterranean Italian buffaloes, through Lactocorder instrument.

8 ORAL ABSTRACTS Cheddar Cheese Nutritional and Therapeutic Delivery Through Use of Nanotechnology

Usman Mir KHAN1, Ammara RIAZ2, Zeliha Selamoglu SELAMOGLU3

1National Institute of Food Science and Technology, University of Agricultre, 38000, Faisalabad , Pakistan 2Department of Zoology, Government College University, Faislabad, Pakistan, Pakistan 3Department of Medical Biology, Faculty of Medicine, Nigde Ömer Halisdemir University, Campus, 51240, Nigde, Turkey, Turkey [email protected], [email protected], [email protected]

Nanotechnologies in food processing and packaging with special attention to their reflection on food quality and safety. As nanotechnology now a days can be used for encapsulation of probiotic bacteria which increases their stability and the solubility of valuable when used in dairy products and their effective and controlled delivery to the body is possible.

Methods: Cheddar cheeses was made by use of Bifidobacterium longum, B. animalis subsp. lactis, Lactobacillus casei sps., and Lb. acidophilus sps. were used as an adjunct in the production process with the help of encapsulation technique. Cheeses were ripened at 4°C and 8°C for 24 weeks.

Results: Effects of ripening temperatures and probiotic adjuncts on proteolysis and sensory evaluation of the cheeses were examined. Higher ripening temperature increased the levels of proteolysis in the cheeses. Product of proteolysis and organic acids released during ripening were important for the flavor of Cheddar cheeses. There were positive and significant correlations between the levels of soluble nitrogen, lactic, acetic, and butyric acids, percentage hydrolysis of αs1‐CN and β‐CN to the scores of cheddary flavor (P < 0.05).

Conclusion: Scores for sour‐acid and vinegary flavors were higher in cheeses with the addition of Bifidobacterium sp. or Lb. casei ripened at 8 °C. The scores were positively and significantly correlated to the level of lactic, acetic, and free amino acids in the cheeses (P < 0.05). The results show that both 4°C and 8°C have potential for use in the ripening of probiotic Cheddar cheeses while comparing with non-capsulated probiotics. Thus, it provides more safety for the food processing system. There is an urgent need for regulatory systems capable of managing any risks associated with nano-foods and the use of nanotechnologies in the food industry.

The authors are grateful to Department of Dairy Technology Postgraduate Laboratory of University of Veterinary and Animal Sciences, Pakistan for providing research facilities and their excellent technical assistance.

[1] Bergamini, C. V., Hynes, E. R., & Zalazar, C. A. (2006). Influence of probiotic bacteria on the proteolysis profile of a semi-hard cheese. International Dairy Journal, 16(8), 856–866. [2] Bevilacqua, A. E., & Califano, A. N. (1992). Changes in organic acids during ripening of Port Salut Argentino cheese. Food Chemistry, 43(5), 345–349. [3] Bhowmik, T., & Marth, E. H. (1990). Role of micrococcus and pediococcus species in cheese ripening. Journal of Dairy Science, 73, 859–866. [4] Blanchette, L., Roy, D., Belanger, G., & Gauthier, S. F. (1996). Production of cottage cheese using dressing fermented by Bifidobacteria. Journal of Dairy Science, 79, 8–15. [5] Bouzas, J., Kantt, C. A., Bodyfelt,

9 ORAL ABSTRACTS The Effects of Dam’s Milk Yield and Milk Components on Calf Birth Weight and Some Growth Traits in Anatolian Buffaloes

Ertuğrul KUL1, Özlem TEPE2

1Department of Animal Science, Faculty of Agriculture, University of Kırşehir Ahi Evran, Kırşehir, Turkey 2Department of Animal Science, Faculty of Agriculture, University of Kırşehir Ahi Evran, Turkey [email protected], [email protected]

This study was carried out to determine the dam’s milk yield and quality traits on the calf birth weight and some growth traits of 98 calves of 98 Anatolian buffaloes calving in 2017 and 2018 years in Çorum province. Effect of stage of lactation on daily milk yield (DMY) and milk fat content were significantly important (P<0.05). DMY, lactation milk yield (LMY) and solids-not-fat (SNF) were affected by calving age. Calving season affected the DMY and fat content (P<0.05), but LMY and SNF were not affected. Suckling period and calf sex on DMY, LMY, fat content and SNF were not significantly important. Effect of calving age of dam on the birth weight was significantly important (P<0.05), but not effect on 6-month and 12-month weight. While effect of birth season on the 6-month weight and 12-month weight were significantly important (P<0.05), the birth weight was not affected. Effects of suckling period and calf sex on the birth, 6-month and 12-month weight were not significantly important.

DMY and LMY were divided into three group as <5.50, 5.50-6.50, >6.50 kg and <1100, 1100-1300, >1300 kg. To evaluate by milk fat and SNF, three groups were formed: <7.50, 7.50-8.50, >8.50% and <9.35, 9.35-9.65, >9.65%. The highest 6-month weight was observed in the calves of buffaloes with high DMY (>6.50 kg) (P<0.05). The birth weight and 12-month weight were not affected by DMY of dam. The highest birth weight and 6-month weight were detected in calves of buffaloes with the highest LMY (>1300 kg) (P<0.05). The birth, 6-month and 12-month weights were not affected by milk fat content of dam and SNF. There was a positive (r = 0.331) and statistically significant correlation with birth weight and LMY (P<0.01). Positively and statistically significant correlations were determined between 6-months weight with DMY (r = 0.339) and LSV (r = 0.267) (P <0.01). There were not statistically significant correlations among birth, 6-month and 12-month weight with milk fat content and SNF.

This research was supported financially by Republic of Turkey, Ministry of Agriculture and Foresty, General Directorate of Agricultural Research and Policies. This research was carried out from a part of M.Sc. thesis of the Özlem TEPE. We would like to thank Mehmet Ali YILMAZ and Çorum Water Buffalo Breeders’ Association for valuable technical assistance.

10 ORAL ABSTRACTS A SNP At the Exon 1 of The Lipoprotein Lipase (LPL) Gene Is Associated with Milk Traits in The Mediterranean River Buffalo

Alfredo PAUCIULLO1, Giustino GASPA1, Giulia CHEMELLO1, Marco IANNACCONE2, Liliana DI STASIO2, Gianfranco COSENZA2

1Department of Agriculture, Forest and Food Sciences, University of Torino, Grugliasco , Italy 2Department of Agriculture, University of Naples Federico II, Portici , Italy [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

Lipoprotein lipase is a key enzyme for the lipid metabolism playing a fundamental role in the composition of fat in adipose tissue as well as in milk. LPL gene was poorly investigated in dairy ruminants, and it has been never studied in river buffalo. Aims of this work were to explore the genetic diversity of LPL (from exon 1 to 4) in this species; to characterize the gene promoter; and to achieve an association study with milk traits. Genomic DNA of 14 individual samples of Mediterranean Italian breed was amplified and sequenced. Thirteen consensus sequences for transcription factors were found. C/EBPα and Oct-1 are the main regulatory motifs together with LP-α, although a deeper investigation is necessary to elucidate the role of the latter element, which in buffalo (and ruminants in general) is lacking the downstream cis-acting motifs (LP-β) compared to human. A total of 11 SNPs have been detected. The SNP g.107A>G was the only polymorphism found at exon level, therefore a genotyping was achieved by PCR-RFLP in a population of 523 Mediterranean buffaloes. The allele frequency was 0.63 and 0.37 respectively for the G and A allele. This SNP was significantly associated with n3-PUFA (P<0.03; 306 individual milk samples), with the GG genotype showing the highest value (+8.25% and +7.14% compared with AA and AG, respectively). The AA and GG genotypes showed higher values also for the milk yield compared with AG, but the estimated difference only approached the level of significance (P=0.07). This study is among the first reports that show an association between milk fatty acids and LPL genotype in ruminants, and it adds further knowledge in the study of genes involved in the regulation of milk fatty acids composition in river buffalo.

11 ORAL ABSTRACTS Welfare Evaluation in Buffalo Species by Risk Assessment Methodology, A Part of The Classyfarm System

Domenico VECCHIO1, Esterina DE CARLO2, Francesca FUSI3, Valentina LORENZI3, Guseppe DE ROSA4, Fabio NAPOLITANO5, Giorgio GALIERO6, Luigi BERTOCCHI3

1National Reference Centre on Water Buffalo Farming and Productions Hygiene and Technologies IZSM, Portici, Italy 2National Reference Centre on Water Buffalo Farming and Productions Hygiene and Technologies IZSM, Portici, Italy 3Centro di referenza nazionale per il benessere animale Istituto Zooprofilattico Sperimentale della Lombardia ed Emilia Romagna , Italy 4Dipartimento di Agraria, Università degli Studi di Napoli Federico II, Portici, Naples,, Italy 5Scuola di Scienze Agrarie, Forestali, Alimentari ed Ambientali, Università degli Studi della Basilicata, Potenza, Italy 6 Istituto Zooprofilattico Sperimentale del Mezzogiorno IZSM Portici, Italy [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

ClassyFarm is an integrated system aiming at the categorization of the farms according to a risk assessment methodology (RA). It is an Italian innovation that allows to facilitate and improve the collaboration and the dialogue between the breeders and the competent authority, in order to raise the level of safety and quality of food of animal origin.

ClassyFarm collects, gather and process data, referred to the following evaluation areas: biosecurity, animal welfare, health and production parameters, animal nutrition, antimicrobial usage, injuries detected at slaughterhouse. ClassyFarm can be applied to several livestock species, included buffalo farms. On the request of the Italian Ministry of Health, CreNbuf in collaboration with CReNBa, have develop a checklist (CL) for the animal welfare assessment in buffalo species, based on RA and included in the ClassyFarm system.

The multiple-choice CL consists of 80 Items. Each Item is scored according to 3 choice categories: “insufficient”, “acceptable” and “excellent”. The assessment system includes non-animal based (N-ABMs) and animal-based measures (ABMs). N-ABMs are divided into 2 macro-areas: Area A (28 Items) “Management factors”; Area B (30 Items) “Housing factors”. ABMs are assessed in Area C (14 Items). The CL has been tested in 341 farms, with an average of 353 head (min10, max 2240). The average overall welfare value was 61.27% (on a scale of 0 to 100%). The values of the specific areas were: Area A 60.20%; Area B 41.34%; Area C 71.55%. At least one potential legislative non-compliance was recorded in 39.8% of the farms.

These CL represent a functional and smart instrument to assign an animal welfare index to each farm, and to provide to veterinarians and breeders a tool for improving farm management and housing conditions by using the data collected in each Area, while respecting farm sustainability, giving answers to consumers and add value to farmers’ correct activities. The Italian Ministry of Health is promoting this system, unique in its kind, also in the European and international context, raising a wide and strong interest.

This research was supported by Italian Ministry of Health, (number project cup: E52I14001190001)

[1] Bertocchi L., Fusi F. (2014). “Guidelines for the assessment of welfare and biosecurity in dairy cattle in loose housing systems”. Brescia, Italy: IZSLER”. [2] Bertocchi, L., Fusi, F., Angelucci, A., Bolzoni, L., Pongolini, S., Strano, R.M., Ginestreti, J., Riuzzi, G., Moroni, P., Lorenzi, V., 2018. Characterization of hazards, welfare promoters and animal-based measures for the welfare assessment of dairy cows: Elicitation of expert opinion. Prev. Vet. Med. https://doi.org/10.1016/j.prevetmed.2017.11.023 [3] De Rosa G., Grasso F., Winckler C., Bilancione A., Pacelli C., Masucci F., Napolitano F., 2015. Application of the Welfare Quality protocol to dairy buffalo farms: Prevalence and reliability of selected measures. J Dairy Sci. 2015 Oct; 98(10):6886-96. doi: 10.3168/jds.2015-9350. Epub 2015 Jul 29. [4] EFSA Panel on Animal Health and Welfare (AHAW); Guidance on risk assessment for animal welfare. EFSA Journal 2012; 10(1):2513. [5] Vecchio D; Tamaio J.E., Bertocchi L; De Carlo E. 2017. Development of a new approach to evaluate Buffalo Welfare, the Ruminant Welfare® project. XVII Latin American Congress Buiatrics, Medellin, Colombia from 1st to 3rd November 2017. Invited lecture

12 ORAL ABSTRACTS Development of Buffalo Production In Mexico

Ismael CORONEL

AMEXBU, Mexico [email protected]

Since the foundation of AMEXBU in 2008, the aim of the organization has been to help producers through the promotion association, identification and solution of problems for development of Mexican buffalo production systems. The aim of this paper is to present from the perspective of the Association, the evolution of buffalo production in Mexico, especially in the last 10 years. Thanks to the effort of Maitret family 3120 Buffaloes arrived to Mexico between 1996 and 1998, through the border with Belize, that were distributed in the states of Campeche and Chiapas, to take advantages of the specie for production in regions of the country with low and flooded areas that additionally produce low quality fodder, After that animals move to other zones of the country such us Campeche, Tabasco, Chiapas, Oaxaca and Veracruz, sites where they have shown their adaptability and superiority over cattle, today it is spread across half of the country’s 32 states, additionally Puebla, Hidalgo, State Mexico, Queretaro, Aguas Calientes, Zacatecas, Nayarit.Today It is estimated that there are around 45,000 buffaloes, with an annual increase of 18%. Its birth rate is 95%, birth weight is in the range of 35 to 45 kg, and 15 months weight is 340 to 360 kg. Most of the farms are dedicated to the production of meat, few farms milk, some of the with mechanical milking. Production systems have different levels of technification, from purely extensive to housing in barns, some breeders have applied reproductive biotechnologies such as artificial insemination. AMEXBU is the natural response of Mexican breeders for the future, especially for the management of the information in all technical issues: health, reproduction and genetic improvement. Also to serve as a bridge between producers and the authorities for the marketing and promotion of the activity, especially safety and sustainability. Big challenges and developments come , associate the largest number of breeders in the country, update a census that allows defining policies and programs to position in the best way Mexican buffalo products

AMEXBU

13 ORAL ABSTRACTS Maize Grain Grown on Mycorrhized Soil Does Not Affect the Rumen of Buffaloes or Dairy Cows

Antonella CHIARIOTTI1, Joan EDWARDS2, Hauke SMIDT2, Gennaro CATILLO1, Luca BUTTAZZONI1

1CREA Research Center for Animal Production and Aquaculture, via Salaria 31, 00015 Monterotondo, Italy 2Laboratory of Microbiology, Wageningen University & Research, Stippeneng 4, 6708 WE, Wageningen, Netherlands [email protected], [email protected], [email protected], [email protected], luca. [email protected]

Arbuscular mycorrhizal fungi (AMF) have mutualistic associations between plants and soil fungi. This benefits plants by improving soil fertility and increasing phosphate absorption, which results in decreased fertilizer requirements. Other reported benefits include improved resistance to pests and pathogens, water stress and salts (Ranganathswamy et al. 2019; Wu et al., 2016). Combined, these benefits decrease production costs and environmental pollution risk. Inoculation of maize seed with AMF does not affect the quality of the forage subsequently produced. However, it has been reported that it can influence the composition of the resulting silage in terms of increased biomass, protein and dry matter (Uzun et al. 2018). Whether this influences the ruminal fermentation of the silage is not clear. It is also not known if AMF treatment results in changes in maize grain that can affect ruminal fermentation. In this study, we investigated the effect of maize grain from an AMF treated crop on the rumen fermentation and microbiota of buffaloes and Holstein Fresian dairy cows.

Maize grains produced from mycorrhized (E) and non mycorrhized (C) crops were fed to six rumen cannulated buffalo and five rumen cannulated Holstein Fresian dairy cows in a cross-over design. The diet of the lactating animals was amended by the inclusion of 5 kg of the C or E maize grain. Rumen content was collected before feeding for analysis of VFAs, pH and microbiota. Microbiota analysis included protozoa direct counts and quantitative PCR of total bacteria, anaerobic fungi and archaea. Statistical analysis was performed using an ANOVA model (SAS, 2001).

Concentrations of total VFA, acetate, propionate, butyrate were only affected by breed (P >0.01), with buffalo having higher values than dairy cows. No effects were found on pH, protozoa direct counts and total bacteria, archaea and anaerobic fungal concentrations (P>0.05). These findings indicate feeding ruminants’ maize grains from AMF treated crops has no detrimental impact on ruminal fermentation or microbiota.

This work was supported by the Italian Ministry of Agriculture (MiPAAF) under the AGROENER project (D.D. n. 26329, 1 april 2016) - http://agroener.crea.gov.it/

[1] Genovese, A.; Marrazzo, A.; De Luca, L.; Romano, R.; Manzo, N.; Masucci, F.; Di Francia, A.; Sacchi, R. Volatile Organic Compound and Fatty Acid Profile of Milk from Cows and Buffaloes Fed Mycorrhizal or Nonmycorrhizal Ensiled Forage. Molecules 2019, 24, 1616 [2] Ranganathswamy M., Kadam G.L., Jhala Y.K. (2019) An Insight into Mycorrhiza Involved in Building Soil and Plant Health. In: Panpatte D., Jhala Y. (eds) Soil Fertility Management for Sustainable Development. Springer, Singapore [3] Uzun, P.; Masucci, F.; Serrapica, F.; Varricchio, M.L.; Pacelli, C.; Claps, S.; Di Francia, A. Use of mycorrhizal inoculum under low fertilizer application: Effects on forage yield, milk production, and energetic and economic efficiency. J. Agric. Sci. 2018, 156, 127–135. [4] Wu, Q.S.; Liu, C.Y.; Zhang, D.J.; Zou, Y.N.; He, X.H.; Wu, Q.H. Mycorrhiza alters the profile of root hairs in trifoliate orange. Mycorrhiza 2016, 26, 237–247.

14 ORAL ABSTRACTS Examining Variability Between Buffalo Farms in Nepal Based on Production Efficiency

Shailesh SHRESTHA1, Cesar REVOREDO1, Rabin RAUT2, Rajendra TIWARI2, Bhuminand DEVKOTA2

1Scotland’s Rural College, United Kingdom 2Agriculture and Forestry University, Nepal [email protected], [email protected], [email protected], [email protected], [email protected]

Nepal has the highest density of buffalo per capita but the lowest productivity in South Asia, a major buffalo production region of the world. The average annual milk yield per buffalo on a farm is 900 litres which is less than half of average yield on farms in India and Pakistan, the other two main buffalo producing countries in the region. There is a wide variability in production efficiency among buffalo farms. This paper looks at the variability in production efficiency between buffalo farms and identifies constraints that renders low efficiency. A farm level survey was conducted with 50 randomly selected buffalo farms in Chitwan region. The survey used a questionnaire to collect farm level physical, management and production information as well as farmers’ intentions and perceptions of buffalo production. A K-means clustering technique was used to group the sampled farms in four clusters - low efficiency; medium low efficiency; medium high efficiency and high efficiency farm groups based on farm size, level of production and input-output score. Production efficiency was highly significant (p<0.01) among these four farm groups with efficiency scores of 0.42, 0.59, 0.73 and 0.88 respectively.A multivariate regression analysis identified production costs; farmer’s education level and share of buffalo income to total household income as the main factors influencing the efficiency among these farm groups. Farms in the lower efficiency farm groups tend to have higher production costs, especially the cost of artificial insemination compared to the farms in higher efficiency group. Other significant characteristics of farms in the low efficiency group are high share of milk for own household consumption, selling milk in local market, less access to dairy co-operatives and receiving low milk price on average. These farms also have a lower share of income from buffalo production to overall household income suggesting a higher reliance on other income sources in comparison to the farms in high efficiency farm group.

The authors are thankful to the Research Directorate at the Agriculture and Forestry University in providing fund to conduct this research work.

15 ORAL ABSTRACTS Determining the Effects of Bypass Fat Supplementation on Milk Yield and Composition in Anatolian Buffaloes

Sezer ÖZ1, Seher KÜÇÜKERSAN2

1TAGEM, Turkey 2Ankara Üniversitesi, Turkey [email protected], [email protected]

A total of 21 early lactating buffaloes allocated three treatment groups according to average milk yield, lactation length, lactation number, live weight and age in this trial. Trial was conducted according to crossover design. No additives supplemented to control group in every period, while 250 g/head palm by-pass fat and animal originated by-pass fat were added to other groups daily ration. The trial lasted 60 days in total for 3 periods, and every period included 15 days of adaptation period, and 5 days of milk sampling.

As a result of the trial, milk yield and feed corrected milk were found statistically non-significant (P>0,05) among all groups. In addition to this milk fat, solid not fat, protein, lactose and its yields as well as milk energy yield were found non-significant (P>0,05). However, higher feed efficiency, milk yield, milk fat, protein and lactose were obtained by animal originated by- pass fat supplemented group than palm by-pass fat supplemented group and control group. Milk pH, temperature, density, freezing point, mineral matter ratio, organic matter ratio, and milk urea-nitrogen were also found statistically non-significant (P>0,05).

As a result, it is determined that animal originated by-pass fat and palm by-pass fat may be added to buffalo rations 250 g/ day per head to increase the milk yield and components without any negative effect. When we compare the groups of which by-pass fat sources are added, it is concluded that animal originated by-pass fat has more positive effects on milk components, and parameters than palm by-pass fat.

[1] AFZAL M, ANWAR M ve MIRZA MA (2007). Some Factors Affecting Milk Yield and Lactation Length in Nili Ravi Buffaloes. Pakistan Veterinary Journal. 27(3): 113-117 [2] AQUINO DL, DEL ROSARIO MV, VERGARA KF ve CRUZ LC (2013). Effects of Augmented Feeding with By-Passed Amino Acid and Slow-Released Non-Protein Nitrogen Supplements on Milk Peak, Lactation Persistency and Post-Partum Reproductive Performance of Brazilian Buffaloes. Buffalo Bulletin, 32(Special Issue 2): 961-965 [3] BARILE VL, TRIPALDI C, PIZZOFERRATO L, PACELLI C, PALOCCI G, ALLEGRINI S, MASCHIO M, MATTERA M, MANZI P, BORGHESE A. (2007). Effects of Different Diets on Milk Yield and Quality of Lactating Buffaloes: Maize Versus Sorghum Silage. Italian Journal of Animal Science, 6 (Suppl. 2): 520-523 [4] BARTOCCI S,TERRAMOCCIA S ve TRIPALDI C(2006). The Utilisation of a High Level Energy/Protein Diet for Lactating Mediterranean Buffaloes: Intake Capacity and Effects on Quanti–Qualitative Milk Parameters. Livestock Science, 99(1-2): 211– 219 [5] BORGHESE A (2005). Buffalo Production and Research. FAO Ed. REU Technical Series. 67: (1-315) [6] BORGHESE A (2010). Development and Perspective of Buffalo and Buffalo Market in Europe and Near East. Revista Veterinaria, 21:20-31 [7] BORGHESE A (2013). Buffalo Livestock and Products. Italy: Rome [8] GARG MR, SHERASIA PL ve BHANDERI BM (2012).Effect of Supplementing Bypass Fat with and without Rumen Protected Choline Chloride on Milk Yield and Serum Lipid Profile in Jaffarabadi Buffaloes. Buffalo Bulletin, 31(2): 91-97 [9] PASHA T (2013). Prospect of Nutrition and Feeding for Sustainable Buffalo Production. Buffalo Bulletin, 32(Spec. 1): 91-110 [10] SHARMA P, GUPTA RS ve BAGHEL RPS (2007). Energy Requirements of Lactating Murrah Buffaloes (Bubalus Bubalis). Buffalo Bulletin,26(4):134-137 [11] ZICARELLI L (1999). Nutrition in Dairy Buffaloes. In: Tionhati, H., Barnabe, V. H., Baruselli, P.S. (Eds.), Perspectives of buffalo husbandry in Brazil and Latin America. Funep, Jabutical: 157-178

16 ORAL ABSTRACTS Utilization of Ensiled Kinnow Waste and Empty Pea Pods as Feed for Buffalo Calves

Manju WADHWA, Japal Singh HUNDAL, Mohinder Pal Singh BAKSHI

Department of Animal Nutrition Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana-141004, India [email protected], [email protected], [email protected]

The left over material (peels, seeds and residual pulp) after extraction of juice from kinnow mandarin (Citrus reticulata) called kinnow waste (KW) constitutes 50% of the intact kinnows. Likewise after shelling peas (Pisum sativum), the empty pea pods (EPP) constites about 55% of intact pea pods. Fresh KW (20% DM) and EPPs (14% DM) were mixed with wheat straw (WS; 94% DM) separately in 75:25 and 80:20 ratio on fresh weight basis to obtain 34-35% DM and ensiled in 10-12 feet long low density polyethylene tubes of 6 feet diameter for 42 days. The ensiled wastes were evaluated by in vitro gas production technique. A 45 days feeding trial was conducted on 12 male buffaloes (Live weight 269.4 ± 7.60 kg) divided in to 3 equal groups to see the effect of feeding ensiled KW-WS or ensiled EPP-WS on nutrient utilization. In the control group the TMR contained concentrate mixture, green fodder and wheat straw in 35:19.5:45.5 ratio on DM basis; while experimental groups contained concentrate mixture, green fodder, wheat straw and ensiled KW-WS or ensiled EPP-WS in 35:19.5:20.5:25 ratio on DM basis. The data of in–vitro and in–vivo studies were analyzed by one way ANOVA.

The in-vitro studies revealed that the true OM digestibility, ME content, volatile fatty acid (VFAs) production were higher (P<0.01) in KW-WS silage in comparison to EPP-WS silage. The methane production was lower (P<0.01) in KW-WS silage, resulting in higher fermentation efficiency in comparison to EPP-WS silage. The daily DM intake in buffaloes was improved (P<0.05) in EPP-WS silage-TMR as compared to control group, but comparable with KW-WS silage based TMR. The digestibility of nutrients was improved (P>0.05) in silage based TMRs as compared to control TMR. The feeding of silage based TMR did not show any affect on blood profile and excretion of total purine derivatives in the urine. The N-retention was improved (P>0.05) in both silage based TMRs as compared to control group. It was concluded that both KW and EPP can be ensiled successfully and effectively utilized in the total mixed ration of buffaloes RKVY (Rashtriya Krishi Vikas Yojana)

17 ORAL ABSTRACTS Evaluation of Anti Mullerian Hormone in Heifers, Normo-ovulatory And Anovulatory Buffaloes

Prakash KEKAN, Shailesh INGOLE, Anagha NAGVEKAR, Simin BHARUCHA, Shambhudeo KHARDE

Mumbai Veterinary College, India [email protected], [email protected], [email protected], [email protected], [email protected]

AMH produced by granulosa cells is an indirect measure of ovarian reserve, and can be a reliable endocrine marker of small antral gonadotropin responsive follicles and ovarian ageing in cows. AMH and AFC was evaluated in the heifers, cyclic, anestrus and repeat breeder Murrah buffaloes, progesterone in cyclic buffaloes and ovarian histopathology of cyclic and anestrus buffaloes. In cyclic buffaloes the mean AFC of 3-5, 5-8 and total AFC >3 mm did not differ significantly whereas AFC >8 mm (P<0.05) and progesterone concentration differed significantly (P<0.01) throughout estrous cycle. No significant difference in mean of AMH levels was observed. But highly significant (P<0.01) individual variation in mean AFC, total follicle counts and AMH levels were recorded during estrous cycle. A positive association between AFC, AMH and fertility in Murrah buffaloes was observed which is due to the capacity of granulosa cell to produce AMH which in turn is dependent on concentration of FSH. A nonsignificant difference in AMH levels of heifer buffaloes was observed during fortnightly collection, whereas, highly significant (P<0.01) individual variation was observed in AMH level of heifers before puberty. The AMH concentration in heifers above 200 pg/ml were pregnant, while below 200 pg/ml were non pregnant, indicating suboptimal fertility in low AMH heifers. A low non significant AMH concentration was observed in anestrus buffaloes. The individual AMH concentration in anestrus and repeat breeding buffaloes differed significantly (P<0.01). The anestrus buffaloes having AMH concentration above 200 pg/ml showed estrus on correcting various factors. The histopathology of ovaries showed significantly high growing follicles in cyclic than anestrus buffaloes. Thus its concluded that there’s a positive association between AFC, AMH and fertility and determination of AMH concentration can be a tool to predict longevity and fertility of dairy animals. The AMH concentration of 200 pg/ml can be cut-off value for Murrah buffaloes to predict the future potential of heifers and young adult buffaloes.

[1] Fallat ME, Siow Y, Marra M, Cook C, Carrillo A. Mullerian-inhibiting substance in follicular fluid and serum: a comparison of patients with tubal factor infertility, polycystic ovary syndrome, and endometriosis. Fertil Steril 1997; 67:962–965. [2] Jimenez-Krassel F, Scheetz D, Neuder L, Ireland J, Pursley J, Smith G, Tempelman R, Ferris T, Roudebush W, Mossa F. Concentration of anti-mullerian hormone in dairy heifers is positively associated with productive herd life. J Dairy Sci 2015; 98: 3036-3045. [3] Pfeiffer KE, Jury LJ, Larson JE. Determination of anti-mullerian hormone at estrus during a synchronized and a natural bovine estrous cycle. Dom Anim Endocrinol 2014; 46: 58–64. [4] Rico C, Fabre S, Medigue C, Clemente N, Clement F, Bontoux M, et al. Anti-Mullerian hormone is an endocrine marker of ovarian gonadotropinresponsive follicles and can help to predict superovulatory responses in the cow. Biol Reprod 2009; 80:50–59.

18 ORAL ABSTRACTS National Project About Anatolian Buffalo Breeding in Turkey

Yusuf KAPLAN, Ali AYAR, Kürşat ALKOYAK, Özden SARIKAYA

General Directorate of Agricultural Research and Policy, Republic of Turkey Ministry of Agriculture and Forestry, Turkey [email protected], [email protected], [email protected], [email protected]

In the world, Turkey that has animal and plant diversity in terms of genetic resources is one of the important countries. Beside of genetic diversification, Turkey’s animal population is significant and it consists of approximately 35.2 million sheep, 10.9 million goats, 17 million cattle and 178.3 thousand Anatolian Buffalo (AB). In parallel with the demand of consumer and increasing buffalo products, Turkey’s buffalo population, which is a decreasing trend until 2011, has started to increase. To improve the AB production, their breeders’ economic and social situation and to develop AB products, Turkish Ministry of Agriculture and Forestry (MAF) has started A National Project about Anatolian Buffalo Breeding in 2011. The project has embraced a multi-stakeholder approach with different partners such as Universities, Research Institutes and Turkey Water Buffalo Breeders Association. In this project, improving AB milk and meat yield, the consciousness of AB breeders and developing breeder association infrastructure are purposed. In five-year periods, the first stage of the carried out the project resulted between 2011 and 2016. The second stage of the project is going to be continued between 2016 and 2021 In accordance with the 2018 year results of the Breeding Project made activities are in 18 provinces on 3.206 farms with 30.000 adult AB actively.

The aim of this study is to demonstrate how to implement the project on farm condition and an overview of general results in Turkey’s buffalo production.

19 ORAL ABSTRACTS Whole-genome Resequencing Reveals Signatures of Selection for Body Size In Chinese Buffalo

Yi ZHANG, Xiangyue MENG, Jingfang SI, Dong LIANG

China Agricultural University, China [email protected], [email protected], [email protected], [email protected]

The Asian water buffalo (Bubalus bubalis) is found on all five continents, with a global population of some 202 million. The recent availability of a high-quality reference genome and of a density genomic marker array for genotyping has triggered a number of genome-wide investigations on diversity, evolutionary history, production traits and functional elements. Chinese buffalo is an important livestock species supplying meat, working power as well as milk in the south of China. Twenty-six local Chinese buffalo populations have been characterized with significant differences in body size. However, genetic mechanism of the growth trait has not been well investigated. In this study, based on genome sequence data, genome scan of selection signatures identified a number of genomic regions that contain putative candidate genes for body size in buffalo. Several strong candidate genes are overlapped with stature genes in cattle and human, suggesting that there are common genes regulating body size in mammals. These findings are valuable for uncovering the gene regulation mechanism of growth traits in buffalo.

20 ORAL ABSTRACTS Potential of Water Buffalo Breeding In Diyarbakır Province

Hasan Hüseyin IPÇAK1, Muzaffer DENLI 2, Muhittin TUTKUN3

1Department of Animal Science, Faculty of Agriculture, Dicle University, Turkey 2 Dicle üniversitesi, Ziraat Fakültesi, Zootekni Bölümü, Turkey 3Dicle üniversitesi, Ziraat Fakültesi, Zootekni Bölümü, Turkey [email protected], [email protected], [email protected]

The aim of this study is to examine the current status of water buffalo breeding and socio-economic structures in the province of Diyarbakır and to provide preliminary information on new investment or project ideas in the region in order to increase the share of water buffalo products in animal production. Located in the Tigris River basin, Diyarbakır has an important strategic position in terms of water buffalo breeding, which is mainly based on wetlands and marshes. Indeed, Diyarbakır ranks the 2nd province in Turkey in terms of the water buffalo population. Moreover, it is the province with the highest number of enterprises registered with the Water Buffalo Breeders Association in Turkey.

As a result of the preliminary feasibility study, it was observed that water buffalo breeding was generally conducted as small family businesses with a capacity of 1-5 heads. The majority of breeders perform a mixed water buffalo breeding with other cattle, and families provide the labor force. Generally, it is reported that water buffaloes are grazed in pastures for 8 months of the year without intensive feed supplementation, that they are fed with poor quality roughage, and that a rootstock water buffalo gives 4-5 liters of milk per day during the lactation period. On the other hand, low-income breeders often use the milk they supply to make yogurt, cheese, butter, or clotted cream to meet their needs, and they sell a small amount on their own. Therefore, the support given to the livestock sector in recent years and the consumer demand, especially for organic livestock and organic products have increased the interest in water buffalo products causing water buffalo breeding to gain importance again. For this purpose, in this region which is advantageous in terms of water buffalo breeding, the share of water buffalo products in animal production can be increased by continuing breeding activities, developing production models suitable for different economic and environmental conditions, solving feed and marketing problems, thus it can contribute to the economy and to raising healthy individuals.

21 ORAL ABSTRACTS Standardization of Environmental Factor Effecting Production Traits by Least Squares Methods an Examples of Application By Excell (lSM-EX)

Şükrü DOĞAN1, Ahmet PEKGÖR2, M. Ihsan SOYSAL3

1Bahri Dağdaş Uluslararası Tarımsal Araştırma Enstitüsü, Turkey 2Necmettin Erbakan Üniversitesi, Fen Fakültesi, İstatistik Bölümü, Turkey 3Namık Kemal Üniversitesi, Ziraat Fakültesi, Zootekni Bölümü, Turkey [email protected], [email protected], [email protected]

In summary, animal breeding relates to intentional selection by humans based on animal performance in a certain environment for predefined and heritable traits. The animals that are superior for desired traits (or combination of traits) related to performance will be selected as breeding animals. Mass selection The most basic way is to rank the animals according to their phenotype and select the best ones for breeding. This method is also called Mass Selection, or Selection on Own Performance. the largest as parents for the next generation. The challenge in selective breeding is to find the best animals as parents for the next generation. Collected information on the performance of the animals, of their pedigree, and of their relatives, need to be evaluated for choosing the best geneticaly superior animals from their contemporaries. There are many methods for ranking the animals according to their genetic merits. Some traits of farm animals In statistical term called discrete variables, recorded in a limited number of classes.Many traits of animals are polygenic, caused by effects of many genes. Many polygenic traits are quantitative and continuous and can be measured in metric units. The variation among animals is represented by the phenotypic variation. The result of a measurement on an animal is called a phenotype.Phenotypic Variation of the animals, is not only determined by the genetic potential, but also by environmental influences. The phenotypic variation observed as consequence of all the genetic and environmental influences and interactions affecting the trait, including errors in measurements can be seperated into componets such as genetic and environmental components. In order to most simply rank the animals according to their estimated genetic merit potentials need to express their standardized phonotypic value relative to the average animal in a population. An expression of an animals performance for a particular trait relative to the herd or contemporary group average is called trait ratio.It is usually calculated for most traits as: [(indiviual record / Average of animals in group) * 100]. So it expresses an estimate of how much better than average an animal is. Mass selection is the most elementary type of selection: based on observed phenotypes. Presented examples of application programed is able to standardize and adjust the data for the magnitudes of these effects were estimated by Least Squares Procedure and linear regression method .Collected quantitavie traits according to the some of sources of variation for 10 different discrete environmental factor and 1 continious factor according to the least squares methods using Excel which is a spreadsheet developed by Microsoft for Windows, It features calculation, graphing tools, pivot tables, and a macro programming language called Visual Basic for Applications.

22 ORAL ABSTRACTS Parameters of Lactation Shape as Related to Days-in-milk In Buffaloes from Two Different Farming Systems

Pencho PENCHEV1, Yordanka ILIEVA1, Tatyana IVANOVA1, Georgi NONCHEV2

1Agricultural Institute – Shumen, Bulgaria 2Agricultural Institute - Shumen, Bulgaria, Bulgaria [email protected], [email protected], [email protected], [email protected]

The aim was to study lactation curve and persistency as related to days-in-milk (DIM) in buffalo cows from two farms – intensive (Fm1) and pasture (Fm2) system with respectively 438 and 330 normal (210-plus days) and 115 and 58 short lactations. Persistency index was computed as the average change in milk yield from 1st to 8th month (PI1) and also as the average decline after established peak month (PIP). ANOVA (LSMLMW and MIXMDL, Harvey 1990) was carried out per each 10-day period (“tenday”) of lactation to outline the lactation curve and to study the effects. The resulted curves show peak yield averagely at 2nd tenday in both farms. Compared to the buffaloes on pasture, those on Fm1 have significantly lower milk in the initial three and in 15th to 21st tendays, the decline in their lactations being slower up to 12th tenday and faster after that. ANOVA’s show greatest effect of parity on milk yield in all tendays as well as of season of calving, and of year on 2nd-12th tenday. The regression of DIM on milk yield is rather strong after 3rd tenday (P<0.001). Persistency (PI1) is affected by year and season of calving (P<0.05) and especially by peak month and DIM (P<0.001). There is a non- significant difference in PI1 in favour of Fm2 (0.940) as compared to Fm1 (0.932). PIP is respctively 0.893 and 0.880. The percentage of lactations shorter than 210 days is as high as 17.8%. The curves are gradual and the shorter the lactation, the lower the persistency – from 0.831 to 0.716; and the lower the peak yield – by 17 to 32% worse than in normal lactation. Long (306-365 days) and very long lactations have higher persistency (respectively 0.950 and 0.955). Noteworthy is that in the lactations over 365 days, the peak is at 4th tenday in which milk yield is by 10% lower than in average normal lactation.

23 ORAL ABSTRACTS Genome-wide Diversity and Population Structure of Indian Swamp and Riverine Buffaloes

D. Ravi KUMAR1, Jayakumar SIVALINGAM2, R S KATARIA2, M Joel DEVADASAN 1, T SURYA1, M R VINEETH 1, Anjali CHOUDHARY1, S K NIRANJAN 2, Archana VERMA1

1ICAR-National Dairy Research Institute, Karnal, Haryana, India 2ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana, India [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

India is the only sub-continent enriched with both riverine and swamp buffaloes. However, the diversity studies and breeding programmes are very few in swamp compared to riverine type. So, the present investigation was focused to study the genomic diversity of the Indian swamp buffalo populations with riverine buffalo and indigenous cattle through the reduced representation approach (ddRAD). A total of 17 samples, 3 swamp buffaloes each from Assam, Manipur and Nagaland were analysed and compared with riverine buffalo (N=4) and cattle (N=4) of India. The results of the study revealed a total of 2,29,733 and 2,19,856 high quality SNPs (Read depth 10) along with 16,498 and 15,201 INDELs respectively in the swamp and riverine buffaloes. The heterozygosity (FST) among the swamp buffaloes was found to be 0.11 between Assamese and Manipuri; 0.20 between swamp (Manipuri) and riverine buffaloes; 0.30 between swamp (Manipuri) and cattle. All the three types of swamp buffaloes cluster together and distantly from riverine buffaloes and further apart to cattle. FIS value among swamp buffaloes was 0.02. The observed and expected heterozygosity in all the swamp buffalo populations was found to be 0.254 and 0.221 respectively. Among the swamp buffaloes, the Manipuri swamp buffalo was found to be genetically distinct and closely related to Nagaland; least to Assamese swamp buffaloes. Principal Component Analysis revealed that PC1 and PC2 explained 36.2% and 24.4% of the total variance. PC1 clusters the swamp and riverine buffaloes separately while PC2 clusters the cattle population. The study evidenced the genomic diversity in the swamp buffalo populations and its uniqueness on comparison with riverine buffalo and cattle.

Authors are Thankful to the Director ICAR-NDRI and the Director ICAR-NBAGR for providing the necessary facilities to carry out this work

24 ORAL ABSTRACTS Determination of Genetic Characterization of Capn1 Gene Exon-1 And -2 And Intron-1 Regions of Anatolian Water Buffalo

Süleyman KÖK1, Selcuk KAPLAN2, Sertaç ATALAY3

1Trakya University, Faculty of Engineering, Department of Genetics and Bioengineering, Turkey 2Namik Kemal University, Faculty of Veterinary Medicine, Department of Genetics, Turkey 3Namık Kemal University, Central Laboratory (NABİLTEM), Turkey [email protected], [email protected], [email protected]

Buffalo CAPN1 geni, kromozom 5’te 22 ekzon ve 21 introndan oluşur. CAPN1 geni, 27146 nükleotit içerir. 23 Anadolu su bufalosu örneği çalışılmış ve sıralama NCBI birincil 3 programı kullanılarak Exon 1 ve 2 ve intron 1 içeren 625 bp buffalo DNA bölgesinde gerçekleştirilmiştir. Anadolu su bufalolarının genotipik karakterizasyonunu belirlemek için CAPN1 geni Exon 1 ve 2 ve intron 1 bölgelerinde spesifik bir SNP olup olmadığı araştırıldı. Genotipleri tanımlamak için sıralama hizalama programı (BioEdit Dizi Hizalama Düzenleyicisi) kullanılmıştır. Anadolu su bufalosu CAPN1 gen sekansında 120044369 inci konum Exon 1 G> T SNP ve 120044661 inci konum intron 1 C> T SNP iki yeni polimorfizm tespit edilmiştir. Normal Anadolu su bufalosunun eksonunda CAPN1 geni, “GGC kodonu” olan 39. Glisin amino asidini kodlar. Bununla birlikte, GGA kodonu mutant Anadolu su bufalosundaki glisin amino asidini de kodlar. Bu nedenle, içinde oluşan SNP’nin protein polimorfizmine yol açmaz. İntron ve ekzon bölgelerinde tespit edilen bu SNP’lerin CAPN1 geninin transkripsiyon düzenleme ve ekleme mekanizması üzerindeki etkileri bilinmemektedir. Bu SNP’lerin Anadolu bufalolarının et kalitesi üzerindeki etkisi, fenotipik gözlemlere dayanan genetik çalışmalar ile belirlenmelidir. İntron ve ekzon bölgelerinde tespit edilen bu SNP’lerin CAPN1 geninin transkripsiyon düzenlemesi ve ekleme mekanizması üzerindeki etkileri bilinmemektedir. Bu SNP’lerin Anadolu bufalolarının et kalitesi üzerindeki etkisi, fenotipik gözlemlere dayanan genetik çalışmalar ile belirlenmelidir. İntron ve ekzon bölgelerinde tespit edilen bu SNP’lerin CAPN1 geninin transkripsiyon düzenlemesi ve ekleme mekanizması üzerindeki etkileri bilinmemektedir. Bu SNP’lerin Anadolu bufalolarının et kalitesi üzerindeki etkisi, fenotipik gözlemlere dayanan genetik çalışmalar ile belirlenmelidir.

25 ORAL ABSTRACTS The Comparison of Lactation Curve for Anatolian Water Buffalo Reared In Istanbul

Mehmet Ihsan SOYSAL1, Mehmet AKSEL2, Eser Kemal GÜRCAN1, Emel ÖZKAN ÜNAL1, Serdar GENÇ3

1Tekirdağ Namık Kemal University, Faculty of Agriculture, Department of Animal Science, Turkey 2Istanbul Water Buffalo Breeders Association, Turkey 3Kırşehir Ahi Evran University, Faculty of Agriculture, Department of Agricultural Biotechnology, Turkey [email protected], [email protected], [email protected], [email protected], [email protected]

Biometry of lactation was investigated with different mathematical models for Anatolian water buffalo reared in Istanbul. The data set were obtained as daily milk yield records of water buffalo in present study. The animal material were constituted from different origined as Italy and Turkey. All animals were grouped for lactation number. Wood (Y=atb.e-ct), quadratic (Y=a+bx+cx2), Wilmink (Y=a+be-kt+ct), logarithmic quadratic (Y=β0 + β1 T + β2 T2 + β3 loge(T2)) models were selected from within the models. The average lactation period were calculated 228.21 and 249.49 day and the average lactation milk yield were calculated 1309.1 and 1747.8 kg and this different were found very important as statistically (P<0.01) for water buffalo origined from Turkey and Italy respectively. Similarly, the average daily milk yield were calculated 5.65 and 7.17 kg and this different were found very important as statistically (P<0.01) for water buffalo origined from Turkey and Italy respectively. Determination coefficients (R2) were calculated for Wood, quadratic, Wilmink and logarithmic quadratic models as 0.71-0.70, 0.66-0.64, 0.69-0.69 and 0.72-0.70 respectively for water buffalo origined from Turkey and Italy. At the same time, persistency (S), maximum milk yields (Ymax) and the time of obtaining the milk yield (Tmax) values were calculated for Wood model. These values were found as persistency (S), maximum milk yield (Ymax) and the time of obtaining the milk yield (Tmax) are 6.03 and 6.19, Ymax values are 7.87and 10.35 kg and Tmax values are 31.05 and 35.20 day respectively for water buffalo origined from Turkey and Italy. In this study, daily milk yield records of Anatolian water buffaloes were used in modeling lactation curves with four models and compares origined of animal and lactation number.

Istanbul Water Buffalo Breeders Association

[1] Soysal MI, Gürcan EK (2000). Comparison of the Mathematical Models in Fitting Lactation Curvesfor Black and White Cattle Raised in Tekirdağ and Kırklareli. 51. Annual Meeting of European Association fo

26 ORAL ABSTRACTS Environmental Effects on Birth Weight and Growth of Anatolian Water Buffaloes In Istanbul

Serdar GENÇ1, Mehmet AKSEL2, Emel Ozkan UNAL3, Mehmet Ihsan SOYSAL3, Kemal GÜRCAN3

1Kirsehir Ahi Evran University, Agriculture Faculty, Department of Agricultural Biotechnology, Turkey 2Istanbul Anatolian Buffaloes Breeding Association, Turkey 3Tekirdag Namik Kemal University, Agriculture Faculty, Animal Science Department, Turkey [email protected], [email protected], [email protected], [email protected], [email protected]

The aim of this study was to determine the factors that were affect the birth weight (BW), sixth month live weight (6MLW) and twelve month live weight (12MLW) of Anatolian Buffaloes. The environmental effects of the district (D), calving year (CY), calving month (CM) and sex (S) on these characteristics were investigated. 5082 Anatolian Buffaloes registered to the Istanbul Anatolian Buffalo Breeding Association were used. The mean and standard errors of BW, 6MLW and 12MLW were 34.62 ± 0.11, 115.6 ± 0.47 and 200.0 ± 1.09, respectively. The effects of D, CY, and S on 6MLW were found to be statistically significant (P <0.05), while CM effect was not significant (P> 0.05). The effects of D, CY, CM and S on 6MLW were found significant (P <0.05). For 12MLW, the effects of CY, CM and S were found to be statistically significant (P <0.05), whereas the effect of D were not significant (P> 0.05). When the breeding process is considered, the factors affecting these characteristics should be meticulously determined. Improvements should be made for the purpose of breeding in line with the determined factors. In this study, which factors (D, CY, CM, S) affecting the traits (BW, 6MLWDA, 12MLW) were determined.

27 ORAL ABSTRACTS Effects of Some Environmental Factors on The Some Growth Characteristics and Lactation Milk Yield and Lactation Period of Anatolian Water Buffalo in Balıkesir, Düzce and Kütahya.**

Ebru ERGÜNEŞ BERKİN, Mustafa KÜÇÜKKEBAPÇI, Cüneyt KAPTAN

Republic of Turkey Ministry of Agriculture and Forestry Sheep Breeding Research Institute, Turkey [email protected], [email protected], [email protected]

In this study, some growth and lactation data were analyzed by using the data obtained from the National Project of Anatolian Water Buffalo Breeding in Farm condition in Balıkesir, Düzce and Kütahya between 2013-2017 years. Birth, 6th month and 12th month live weights were evaluated. Year, province and birth season were used as heterogeneity sources and gender were used as co-variable. All heterogeneity sources and co-variable were found statistically significant on interested weights (P<0.001). There was no effect of the first 3 years on 12th month live weight and the increase in the last two years was statistically significant (P <0.001).

5721 lactation data of the same provinces were evaluated. The effect of province, birth season and years on lactation milk yield and lactation length has been evaluated by factorial analysis and results showed that all interactions were not statistically significant. In that case only main effects have been interpreted. 40,61% of births were in spring, 37,39% in summer and 22,01% in other months. The effect of birth season on lactation milk yield and lactation period was statistically significant (P<0.001). In buffaloes giving birth in winter, lactation milk yield is higher and lactation period is longer. In these buffaloes, milk yield at the beginning of lactation is low, but the recovery of pasture with the arrival of spring is one of the factors that increase milk yield. Fluctuations in lactation milk yield and duration have been observed over the years.

When the outputs from this project are examined, it is seen that the farmers have also made positive improvements such as improving the awareness of keeping records, improving the conditions of care and feeding, and rejuvenating the blood to prevent relatives. Thus, besides the increase in the number of buffaloes, important steps have been taken to increase the milk yield, which is the first target of the project. Furthermore, it is clear that further studies are needed to establish other efficiency and performance parameters.

** The authors would like to thank Republic of Turkey Ministry of Agriculture and Forestry and The General Directorate of Agricultural Research because of their support for the National Project of Anatolian Water Buffalo Breeding in Farm condition.

28 ORAL ABSTRACTS Influence of Environment and Some Single Nucleotide Polymorphisms in Growth Hormone Receptor Gene on The Weight of Anatolian Buffalo Calves at 12 Months of Age

Metin ERDOĞAN, Mustafa TEKERLİ, Koray ÇELİKELOĞLU, Özlem HACAN, Serdar KOÇAK, Zehra BOZKURT, Samet ÇİNKAYA, Mustafa DEMİRTAŞ

Afyon Kocatepe University, Turkey [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

The aim of this study was to reveal the effects of three SNPs in 31,601,630 bp ─ 31,602,797 region of Growth Hormone receptor gene (GHR) located on buffalo chromosome 19 and some environmental factors on weights at 12 months of age of 3012 Anatolian water buffaloes. Data were collected from different growing sites of Afyonkarahisar province in the scope of genetic improvement project in public hand. Blood samples were obtained from the buffalo calves of the member breeders of Afyonkarahisar Buffalo Breeders Association. Molecular analysis was realized by the method of Çelikeloğlu et all. (2015). After the completion of processes, data coming from shared recording software “MandaYildizi” (BuffaloStar) and molecular findings matched in Microsoft Excel software and a least squares analyses was performed. ANOVA indicated the SNP in the BBU19:31,601,784 bp of GHR gene (G3678A), growing cite, birth year and season, sex, age of dam and birth weight had significant (P<0.05) effects on the weight of buffalo calves at 12 months. Genotype AA calves were heavier than the genotype GG calves. Genotype AG calves were in the middle of these two genotype. The least squares means for AA, AG and GG genotypes were 168.60±3.12, 165.40±2.59 and 162.63±1.83 kgs respectively. Calf weights showed some gradual increase in years from 153.42±2.14 to 185.04±2.15 kgs. Calves born in autumn and winter were approximately 9 kgs heavier than the others. Female calves were also 5.11 kgs lighter when they reached to 12 months of age. Also, the effect of other two SNPs was not found significant.

It was concluded that the genetic structure of Anatolian water buffaloes represents some opportunities for genomic evaluation and above mentioned environmental factors must be considered when calculating breeding values.

29 ORAL ABSTRACTS Identification of New Novel Single Nucleotide Polymorphisms in Growth Hormone (GH) Gene in Water Buffalo (bubalus Bubalis) Populations In Turkey

Raziye İSIK1, Emel Özkan ÜNAL2, Mehmet Ihsan SOYSAL2

1Faculty of Agriculture, Department of Agricultural Biotechnology, Tekirdağ Namık Kemal University, Tekirdağ , Turkey 22Faculty of Agriculture, Department of Animal Science, Tekirdağ Namık Kemal University, Turkey [email protected], [email protected], [email protected]

Growth hormone (GH) plays a crucial role on physiological regulation such as growth, body composition and carcass traits, reproduction, proliferation of mammary cell and lactogenesis. For this reason, this study was conducted to investigate the growth hormone (somatotropin-like) gene polymorphisms in 150 water buffalo (Bubalus bubalis) from Turkey. 403 bp long partial intron 4, exon 5, 3’ UTR regions of GH gene and 347 bp long exon-intron 3 and partial exon 4 regions of GH gene were amplified and PCR products analyzed via DNA sequencing method. The seven genotypes were identified in 347 bp long region of the GH gene. The four genotypes were identified in 403 bp long region of the GH gene. The partial DNA sequence of the GH gene in water buffalos was reported and these sequences were deposited to NCBI Genbank database. These SNPs may have an effect on economic traits such as body composition and carcass traits, reproduction, milk yield and content in water buffalo populations and use for water buffalo breeding.

30 ORAL ABSTRACTS The Genomic Revolution in Agriculture Research and Breeding

Andre EGGEN

AgriGenomics Manager, Illumina Inc., United States [email protected]

A paradigm shift is taking place with the application of genomics to the design and implementation of livestock breeding programs, promising gains across the value chain. For breeders, breeding organizations and the livestock industry, genomics is expected to increase efficiency and productivity of animal breeding while for consumers and the processing sector it should enhance security and quality of animal products. Genomics proposes further opportunities to improved selection accuracy while decreasing the costs, reducing generation intervals and exploiting new sources of polymorphisms.

This major (r)evolution is greatly facilitated by accessing a reference population with 1. accurate phenotypes and 2. genotypes covering the entire genome to compute a genomic prediction equation.

Individuals of choice to compose the reference population are usually progeny-tested animals as they have reliable phenotypic information from a large group of daughters. However, for small livestock populations or developing countries, the number of progeny-tested animals may be quite limited. Recent studies have demonstrated that the inclusion of female in the reference population would greatly contribute to the increase of reliability of genomic predictions while compared to parent averages or using only males in the reference population (Boison et al. 2017; Su et al., 2016; Thomasen et al., 2014)

Another factor to take into consideration is the density of the SNP array to be used to genotype animals from the reference population and later the candidates for selection. While an intuitive approach would argue “the more the better”, several papers are demonstrating that SNP arrays with very high density (400-800K SNP) are not bringing a cost effective increase in the accuracy of the genomic prediction : for Boison et al. (2017), only a slight increase in reliability (3.3%) in their Gyr (Bos indicus) population was observed when marker density increased from 15,000 to 496,606 markers.

From a pure genomic point of view, the broader adoption of Genomic Selection in small populations and developing countries would therefore benefit having access to cost effective mid to low density SNP arrays that would be optimized to capture the genetic variability in the specific population. Options to developed such SNP arrays will be discussed together with latest development in next generation sequencing that could help setting up a full genomic information pipeline from low density genotypes to full sequence data.

31 ORAL ABSTRACTS A Study on The Diffusion of Innovation Process in Buffalo Owners in Punjab Pakistan

Muhammad Asif YASEEN1, Ahmad MALIK1, Arfan ASGHAR1, Humaira JABEEN2, Mansoor MUBEEN3, Fayyaz AHMAD

1Livestock Experiment Station Chak Katora Hasilpur District Bahawalpur under Buffalo Research Institute Pattoki, Pakistan 2Deputy Director Livestock Hasilpur District Bahawalpur, Pakistan 3LPRI, Bahadurnagar, Pakistan 4LPRI Bahadurnagar, Pakistan [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

Pakistan has a large number of animal populations which contribute about 11.1% of the National GDP and 58.9% of Agricultural contribution towards the National GDP (economic survey of Pakistan 2017-18). Livestock provides insurance to farmers and guarantees to fulfill day to day needs of subsistence farmer’s family. Pakistan possesses twomain buffalo breeds. These breeds are reared for milk and meat purposes.present study was designed to study the effect of new technologies on adoption process in buffalo owners in Punjab. Correlation analysis of the results shows a positive correlation between landholding and number of buffaloes kept (0.379), landholding and number of cows kept (0.741), land holding and FMD vaccination (0.33), between landholding and HS vaccination (0.19) and landholding and deworming (0.27) and landholding and use of Sadabahar seed varieties (0.35). However a non-significant correlation was observed between landholding and AI (0.10), and landholding and health status of the animals. A positive correlation was observed between education level and number of registered farmers (0.37), education and vaccination (0.178), education and deworming (0.15), education and use of Multicut fodder seed (0.38). However, a negative correlation of registered farmers with H.S vaccination (-0.06), deworming (-0.16), FMD vaccination (-0.37) and dipping / spraying of animals (-0.27) was observed in the study.

Glory does not descend upon the people. It is the people who must raise themselves to the level of attaining it. I acknowledge all our colleagues who are involved in serving the humanity with love and fervour.

32 ORAL ABSTRACTS Genetic Evaluation of Nili-Ravi Buffaloes Registered Under the Progeny Testing Program in Punjab, Pakistan

Maqsood AKHTAR1, Muhammad SHAFIQ2, Riaz HUSSAIN 3, Tariq MEHMOOD1, Muhammad IMRAN1, Irem ANWAR1, Muhammad SAJJAD 4

1Buffalo Research Institute Pattoki District Kasur, Pakistan 2Research Centre for Conservation of Indigenous Breeds, Jhang Pakistan, Pakistan 3Department of Livestock and Poultry Production, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan, Pakistan, Pakistan 4University of Agriculture, Faisalabad, Pakistan, Pakistan [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

Nili Ravi Buffalo Progeny testing program (PTP) continues since 1983 in Punjab (Pakistan). Animals are genetically evaluated and ranked for selection of superior bulls and bull mothers. The pedigree and performance recording of Institutional & private herds in the PTP of Nili-Ravi buffaloes is organized and monitored by the two Government institutions “Buffalo Research Institute, Pattoki” & “Research Centre for Conservation of Indigenous Breeds, Jhang” in the L& DD Department Punjab. Present study reports the current genetic evaluation status of Nili-Ravi buffaloes under this program. Data on 31631 lactations of 18372 Nili-Ravi buffaloes sired by 522 bulls (1965 to 2018) in nine different herds (including seven institutional & two private herds) were analyzed using BLUP procedure. A repeatability animal model was employed for the analysis of milk yield records. The year of calving was divided into two seasons (summer and winter) and herd-year- season combinations were defined with at least five observations in any combination. Age at calving was fitted in model as covariable. Lactations were truncated at 305 days as milk yield uptill 305 days was considered as the objective trait. If however, yield information upto 305 days was missing, yields were regressed back to 305 days using models suggested previously (Khan et al., 2012). The accuracy for Estimated Breeding Value (EBV) of every animal was also estimated. The EBVs of 9930 buffaloes were found positive, while, 7919 buffaloes had negative EBVs. The highest EBV among the buffaloes was +1459 kg and lowest EBV was observed as -843.6 kg. Out of 522 sires used during the reported period, 426 were used as AI sires all over the Punjab, while remaining 96 as natural service bulls. The overall EBV of sires ranged from -645.7 to +704.5 kg. For bulls having more than 10 daughters (n=186), positive EBVs ranged from +6.639 to 704.5kg). The accuracy of EBVs for AI bulls ranged between 24 and 97%. Results suggest strengthening of PTP to improve accuracy. Expansion of the PTP to test more bulls will help improve intensity of selection for tangible genetic gain.

33 ORAL ABSTRACTS Effect of Vitamin E and Selenium Administration on Prepartum and Postpartum Performance of Nili Ravi Buffaloes

Usman AYAZ1, Muhammad ARIF1, Irem ANWAR2, Asif YASIN3, Mohsin RAZA4

1Livestock Experiment Station Haroonabad, Pakistan 2Buffalo Research Insitute, Pattoki, Pakistan 3Livestock Experiment Station Chak Katora, Pakistan 4KBCMA College of Veterinary & Animal Sciences, Narowal , Pakistan [email protected], [email protected], [email protected], [email protected], [email protected]

The objective of the present study was to observe the effects of Vitamin E and Selenium (Se) on different productive and reproductive traits of Nili Ravi buffaloes before and after parturition. Twenty multiparous pregnant buffaloes were randomly selected at Livestock Experiment Station Haroonabad, BRI Pattoki and divided in two groups each of 10 animals. Group 1 (n=10) was kept as control with no treatment provided. Group 2 (n=10) was administered with 20 ml of Valenium® (Tocopherol acetate 25 mg/ml, Sodium Selenite 2.2 mg/ml) 30 days and 15 days prior to expected date of calving and then at 30 day interval up to 120 days post calving. All the animals were kept under the similar feeding and management practices. Parameters studied included birth weight of calves (BW) at calving and 120 days Milk production (MP), Number of animals showing estrus, Conception rate (CR), and Body condition score (BCS). Results on reproductive performance showed that Vit. E and Se significantly (P˂0.05) reduced the service period in treated group (80.85±30.53 days)as compared to control group (156.6±92.3 days). 70% of buffaloes showed estrus with CR of 40% in treated group whereas 30% showed estrus with CR of 10% in control group. Results on productive performance showed increase in MP in treated group (10.1 kg/day) as compared to control (9.3 kg/day) but found non-significant at (P˃0.05). BW of calves showed a significant trend (P˂0.05) in treatment group (39±1.94kg) as compared to control (38±1.15kg). BCS of treated group (2.7±0.34) and control group (2.95±0.43) differ non-significantly (P˃0.05). The present study concluded that Vitamin E and Selenium have a strong positive correlation with reproductive performance of Nili Ravi buffaloes in their late gestation period.

34 ORAL ABSTRACTS A Comprehensive Study on Surgical Affections of Teat in Buffalo (bubalus Bubalis) Presented at Surgery Teaching Hospital, University of Veterinary and Animal Sciences, Ravi Campus, Pattoki.

Muhammad Hassan SALEEM1, Ayesha SAFDAR2, Nasir IQBAL2

1University of veterinary and animal sciences, lahore, Pakistan, Pakistan 2university of veterinary and animal sciences, Lahore, Pakistan, Pakistan [email protected], [email protected], [email protected]

Various surgical affections of teats in buffaloes were studied and the order of their frequency was, teat canal fibrosis (n=71; 25.91%), teat orifice blockage (n=53; 19.34%), streak canal fibrosis (n=46; 16.78%), teat laceration (n=44; 16.06%), teat fistula (n=28; 10.21%), udder edema (n=13; 04.74%), relaxed teat sphincter (n=11; 04.01%) and udder fibrosis (n=07; 02.55%). Faulty milking practices were considered as exciting or predisposing factor for many of these conditions. Use of teat instruments like teat bistoury and teat tumor extractor was successfully used for correction of various surgical teat affection. Intra mammary infusions along with surgical corrections were proved effective but resulted in economic losses due to decrease in milk production was observed. In certain cases of teat laceration, local analgesia was also recommended. Delayed cases and non-responding to surgical treatment resulted in permanent drying of affected quarter. WVC UVAS, Chairman Department of CMS

35 ORAL ABSTRACTS Evidence That Cocaine- And Amphetamine-regulated Transcript (cart) Is A Potential Regulator of Follicular Atresia

Chunyan YANG, Haiying ZHENG, Jiaxiang HUANG, Nongqi YU, Jianghua SHANG

Guangxi Buffalo Research Institute, China [email protected], [email protected], [email protected], [email protected], [email protected]

The mechanisms regulating development of a single (dominant) follicle capable of ovulation during each follicular wave in cattle and atresia of remaining follicles (dominant follicle selection) are not well understood. We recently obtained evidence that cocaine- and amphetamine-regulated transcript (CART), a potent anorectic neuropeptide, is expressed in the buffalo ovary and follicles. The objectives of this study was to determine and regulatory role of the buffalo CART on follicular development. The buffalo follicles were divided into big (>8mm), medium (4-6mm) according to the diameters of the follicles, the follicles were isolated from the ovaries. Then, follicle fluid was collected to analyze the progesterone and estradiol concentrations, and the granulosa cells were collected to extract RNA determine the expression of CART mRNA by q-PCR. As a result, amounts of CART mRNA was greater in medium (subordinate) follicles versus big (dominant) follicles (2.37±0.48 vs. 1.28±0.28, p<0.05), and the concentration of estradiol in medium follicles was lower than that of big follicles (391±104 vs. 441±106 pmol/L, p<0.05. ), while the concentration of progesterone has no difference between the medium and big follicels (23.30±1.71 vs. 23.73±0.57 ng/mL, p>0.05), these results indicated a negative association of expression of CART with follicle basal estradiol production by buffalo granulosa cells, while has no effect on the production of progesterone. In conclusion, CART may regulate the health status and follicular astesia according to adjust the production of estradiol by granulosa cells.

This work was supported by the National Key Research and Development Program of China (2017YFE0113800), the Key Research and Development Program of Guangxi China (GuiKeAB1850013) and the National Natural Science Foundation of Guangxi China (2018GXNSFAA050129).

36 ORAL ABSTRACTS An Offspring Delivered by The Transgenic Cloned Buffalo Cow

Jiang-hua SHANG1, Chun-yan YANG1, Hai-ying ZHENG1, Jia-xiang HUANG1, Hua-zhong LIU2

1Buffalo Research Institute, Chinese Academy of Agricultural Sciences, China 2College of Chemistry & Environment, Guangdong Ocean University, China [email protected], [email protected], [email protected], [email protected], [email protected]

Buffalo is an excellent mammary gland bioreactor animal in tropical and subtropical regions with high temperature and humidity. In this study, buffalo transgenic cloned embryos were produced successfully by using the ear skin fibroblast cells stably expressing EGFP via liposome-mediated transfection with the eukaryotic expression vector pEGFP-N1 and G418 selection from a Murrah buffalo cow. The results showed that the fusion, cleavage and blastocyst rates of transgenic cloned embryos were 82.98%, 61.05% and 27.37%, respectively, which were no significant difference in statistics from those of non-transgenic cloned embryos. The frozen-thawn survival rate of transgenic cloned blastocysts was also similar to that of non-transgenic cloned blastocysts. The obtained transgenic cloned blastocysts were 100% in EGFP positive by fluorescence examination and 17 of them were transferred to 12 recipients. Three pregnancies (25.0% pregnancy rate) were found after one-month early pregnancy diagnosis and one healthy female calf was born on August 25, 2014 after full term pregnancy. The calf genome has been integrated with exogenous EGFP after examination of samples from ear skin fibroblasts, cartilage tissue and peripheral blood cells all express the report gene EGFP. This is the first transgenic cloned female calf of Murrah buffalo and is still alive in good condition. In order to prove the reproductive performance and genetic stability of exogenous genes, this transgenic cloned buffalo cows was mated with a Murrah buffalo bull and delivered a 23.0 kg female calf on May 31, 2019, were obtained. The calf is in good health and is the world’s first offspring of transgenic cloned buffalo. The research results indicate that the transgenic cloned buffalo can not only grow up to adulthood, but also get normal pregnancy to deliver the healthy offspring and produce milk, the important raw material of mammary gland bioreactor, which are also laid a solid theoretical and practical foundation for research on mammary gland bioreactor in water buffalo.

This work was supported by the National Key Research and Development Program of China (2017YFE0113800) and the National Natural Science Foundation of Guangxi China (2018GXNSFDA050013).

37 ORAL ABSTRACTS Physicochemical and Textural Characteristics of Sous-vide Cooked Buffalo Beef

Chirasak PHOEMCHALARD1, Tanom TATHONG2, Pitukpol PORNANEK3, Suthipong URIYAPONGSON4

1Department of Agriculture, Mahidol University, Amnat Charoen Campus, Amnat Charoen 37000, Thailand, Thailand 2Department of Food Technology, Faculty of Agriculture and Technology, Nakhon Phanom University, Nakhon Phanom 48000, Thailand, Thailand 3Department of Animal Science, Faculty of Natural Resources, Rajamangala University of Technology Isan, Sakon Nakhon Campus, Thailand 4Department of Animal Science, Faculty of Agriculture, Khon Kaen University, Khon Kaen 40002, Thailand, Thailand [email protected], [email protected], [email protected], [email protected]

This research examined the effects of single step sous vide cooked directly at temperature of 55℃ (SV55) and 59℃ (SV59) for 12 h, multi-step sous vide cooked at first temperature of 55℃ and 59℃ for 6 h and second temperature of 70℃ for 6 h (M55-70 and M59-70), and traditional cooking at 70℃ (TC70) for 45 min on the physicochemical and textural characteristics of longissimus dorsi muscle from swamp buffalo. Meat pH, surface color, browning index (BI), cooking loss (CL), shrinkage, shear force (SF), and texture profile analysis (TPA) were measured. The results obtained indicated that cooked pH was the lowest in TC70. M55-70 and M59-70 samples had much higher CL, longitudinal shrinkage (LS), transversal shrinkage (TS), and BI. Traditional treatments (TC-70) showed higher L*, a*, and C* values; however, total color difference (∆E) was greater in SV55 group. Comparing to other treatments, redness change (∆a) in both M55-70 and M59-70 was more constant. TC70 was also tougher meat as higher SF, hardness, springiness, cohesiveness, gumminess, chewiness, and resilience; while, SV55 presented the most tender. Thus, the sous vide cooked directly at temperature of 55℃ for 12 h had a potential plus to keep up moisture content and physical form and also produce tender meat. However, about color appearance, in sample treated double temperatures gained the best.

38 ORAL ABSTRACTS PLF (Precision Livestock Farming) Technologies Usage for Water Buffalo Breeding

Orhan ERMETIN

Yozgat Bozok Üniversitesi/Ziraat Fakültesi/Zootekni Bölümü, Turkey [email protected]

Water buffalos, a farm animal with an important economic production in the world, are used for meat, milk, traction, hide production and manure. There are approximately 194 million head of buffalos in the world. Ninety seven percent of them are in Asia, 2% in Africa, and 1% in South America, Australia and Europe (FAOSTAT, 2014). The majority of the buffaloes fed for milk are in India, Pakistan, China, Egypt and Nepal. Usually being raised in extensive conditions, compared to other farm animals, the buffalos have many advantages such as better conversion of poor quality roughage to milk and meat, more resistance against to diseases etc.

Ensuring the health and well-being of an animal, achieving good reproduction performance and reducing environmental impacts per animal product are essential for sustainable livestock breeding. The use of PLF (Precision Livestock Farming) information and technology to increase animal health, welfare and farm productivity is a developing scientific and commercial area nowadays. PLF technologies provide a tool for manufacturers by continuously monitoring and controlling environmental, physiological and behavioral variables.

The applications of research and technological advances in livestock farming in the world have initiated the development of precision livestock farming. Through this technology, sensors (cameras, microphones and accelerometers), images, sounds and movements are used to determine the wellbeing of animals without disturbing animals and to prepare algorithms to predict productivity. For early detection, physiological status and well-being of animals, precision livestock technologies allow for remote detection and monitoring of images, sounds, monitoring data, weight, and biological measurements by using the real-time analysis of animals.

This study is carried out to better understand the use of PLF information and technologies in water buffalo breeding through analyzing the relevant applications of PLF information and technologies used for different farm animal breeding.

39 ORAL ABSTRACTS Development of Rapid Multiplex PCR Assays for Identification of Major Mastitis Pathogens in Buffalo’s Raw Milk

Muhammad SAFDAR1, Mehmet OZASLAN2, Rozhgar A. KHAILANY3

1CHOLISTAN UNIVERSITY OF VETERINARY AND ANIMAL SCIENCES, BAHAWALPUR, Pakistan 2GAZIANTEP University, Turkey 3Department of Biology, College of Science, University of Salahaddin, Iraq [email protected], [email protected], [email protected]

Dairy products which are made from the non-pasteurized milk have been responsible for different bacteria outbreaks and must be harmful for consumers.Accurate identification of these species related to mastitis is very necessary for proper treatment and to control this disease.The present study was intended for the simultaneous detection of the Streptococcus uberis (S. uberis), Streptococcus Paraubaeris (S. Paraubaeris), Streptococcus aglactiae (S. aglactiae), Streptococcus dysglactiae (S. dysaglactiae), Staphyloccousaureus (S. aureus) and Escherichia coli (E. coli) extracted from the buffalo’s raw milk samples. A total number of the 150 samples were collected from the different areas of district Muzaffargarh, Punjab, Pakistan. For the bacterial isolation milk samples were cultured on various culture media. The isolated bacteria were identified by performing the simplex and multiplex PCR. By using simplex and multiplex PCR the isolates were identified and obtained PCR products were visualized by the gel electrophoresis. Out of 150 samples collected from different areas of district Muzaffargarh, Punjab, Pakistan, results had shown the presence of S. uberis 32(21%), S. Paraubaeris45(30%), S. aglactiae 29(19%), S. dysaglactiae 30(20%), S. aureus 48(32%), and E. coli 51 (34%). Taken together, all data indicated that the developed multiplex PCR assay was a simple, rapid, sensitive, specific, and cost-effective detection method for all pathogens from buffalo’s raw milk samples.

The authors would like to thank you CUVAS, Bahawalpur and HEC-PAKISTAN for giving us bacterial strains and the necessary facilities.

[1] Amin A S, Hamouda R H and Abdel-All A A. 2011. PCR assays for detecting major pathogens of mastitis in milk samples. World Journal of Dairy and Food Sciences 6: 199-206. [2] Bansal B K and Gupta D K. 2009. Economic analysis of bovine mastitis in India and Punjab—A review. Indian Journal of Dairy Sciences 67: 337-45. [3] Goli M, Ezzatpanah H, Ghavami M, Chamani M and Doosti A. 2012. Prevalence assessment of Staphylococcus aureus and Streptococcus agalactiae by multiplex polymerase chain reaction (M-PCR) in bovine sub-clinical mastitis and their effect on somatic cell count (SCC) in Iranian dairy cows. African Journal of Microbiology Research 6: 3005-10. [4] Muhammad Safdar & Yasmeen Junejo, 2015.A multiplex-conventional PCR assay for bovine, ovine, caprine and fish species identification in feedstuffs: Highly sensitive and specific. Food Control, 50, 190-194.

40 ORAL ABSTRACTS Impact of Length of Lactation on Milk Yield and Calving Interval for The Mediterranean Italian Buffalo Breed

Emanuela PARLATO, Chiara CASO, Angelo COLETTA

RISBUFALA, Italy [email protected], [email protected], [email protected]

The objective of this work was to evaluate effects of length of lactation, in days(LL1: 180-210; LL2:211-264; LL3:265- 275; LL4: 276-300; LL5:>= 301) and calving interval length in, days (CI1: 350-400; CI2: 401-450; CI3: 451-500; CI4: >= 501) on total milk yield (MY) produced in a lactation, to optimize the economic profitability of Mediterranean Italian dairy buffalo breeding during the productive life. Data were obtained from 1.000 Mediterranean Italian buffalo on 1st to 5th parity calved between years 2010 to 2017 on 26 dairy farms of 8 provinces of Italy. Proc GLM of SAS was the procedure used to estimate the MY in LL x CI classes (20 classes). Fixed effect of herd-year, age within parity, and productive level of buffalo was included in the model. The model well fitted the data with a coefficient of determination, R2 = 0.90. MY increases of 263 kg from LL1 (2.875 kg) to LL3 (3.139), whereas, MY for LL3 was not statistically different to MY of LL4 (3.199 kg). LL5 showed the largest MY (3.315 kg). However, the increase of MY from LL3 to LL4 was only of 176 kg. Length of current lactation showed a significant effect on MY per classes of next calving interval length (CI). In fact, for LL1, MY decreased of 235 kg of MY from CI1 (2.926 kg) to CI3 (2.691 kg), whereas, slightly decreased for LL2, LL3 and LL5, from CI1 to CI3 (-44 kg, -54 kg, and -51 kg, respectively. For LL4, MY increases of 6 kg from CI1 to CI3. Optimal LL should be assessed in relation to the increasing of CI in the next lactation. In fact MY was highest for middle LL and CI. The highest individual profitability was reached by buffalo in LL3. For this LL class, buffalo will produce more MY in the current lactation and reduce the CI to obtain the highest MY per CI class.

[1] Pollot, G.E. (2011) Short communication: Do Holstein lactations of varied lengths have different characteristics? Journal of Dairy Science 94, 6173-6180. doi: 10.3168/jds.2011-4467 [2] Van Amburgh, M.E., Galton, D.M., Bauman, D.E., Everett, R.W. (1997): Management and economics of extended calving intervals with use of bovine somatotropin, Livestock Production Science 50, 15-28. doi: 10.1016/S0301-6226(97)00069-9

41 ORAL ABSTRACTS Comparison of Natural and Synthetic Anti-oxidants on Instrumental Color, Oxidative Stability and Sensory Properties of Young Buffalo Meat Burger Patties

Muhammad Hayat JASPAL1, Adeel MANZOOR2, Aneeqa SARDAR3, Muhammad HASSAN2, Sher ALI2, Sohail AHMAD2, Iftikhar Hussain BADAR2

1University of Veterinary and Animal Sciences, Lahore, Pakistan, Pakistan 2University of Veterinary and Animal Sciences, Pakistan, Pakistan 3Livestock and Dairy Development Department., Pakistan [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

In response to consumers’ preference for natural products, effect of natural antioxidant viz. Moringa oleifera Leaves Extract (MLE) and synthetic antioxidant viz. Ascorbic Acid (AA) in improving the instrumental color and oxidative stability as well as their potential to affect sensory attributes of buffalo meat burger patties has been investigated. The MLE was used in 0.3%, 0.5% and 0.7% concentrations while AA was used in 0.01%, 0.05% and 0.1% concentrations. Buffalo lean meat was ground and mixed to achieve approximately 20% fat. Both antioxidants were added at given concentrations and patties were packed in High Oxygen Modified Atmosphere Packaging and stored at 4ºC for 7 days. Instrumental color was recorded throughout 7 days. Thiobarbituric Acid Reactive Substances (TBARS) were measured to determine lipid oxidation at 0, 3 and 6 days of storage. For sensory evaluation, patties were cooked to the core temperature of 74ºC and judged on 8 points hedonic scale at 0, 3 and 6 days. Both MLE and AA significantly (P<0.05) improved instrumental meat color. However, 0.1% AA provided best results followed in order by 0.05% AA, 0.5% MLE and 0.7% MLE. When measured antioxidant activity, 0.1% AA and all concentrations of MLE significantly (P < 0.05) reduced lipid oxidation on day 3, while on day 6, 0.1% AA exhibited maximum antioxidant activity followed by anti-oxidant potential of 0.5% MLE, 0.7% MLE and 0.05% AA. All these three showed similar results. However, sensory panel results did not showed any difference among MLE or AA treated and untreated beef patties in terms of odor, flavor and cooked color. In conclusion, MLE and AA as antioxidants are equally effective in improving instrumental color and oxidative stability of beef patties. Likewise, AA 0.1% and MLE 0.5% concentrations have increased antioxidant activity without effecting sensory attributes of Buffalo meat patties burgers.

42 ORAL ABSTRACTS Efficiency of Fourier Harmonic Analysis in Predicting Fertility in Water Buffalo Bulls

Peregrino G. DURAN1, Danilda HUFANA-DURAN1, Matt Daniel PERALTA1, Emma VENTURINA1, John PARRISH2, Hernando VENTURINA1, Fe VENTURINA1, Felomino MAMUAD1

1Philippine Carabao Center/Central Luzon State University, Philippines 2University of Wisconsin-Madison, United States [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

Laboratory methods to select high fertility semen and eliminate low fertility semen is needed in the water buffalo industry. Fourier harmonic analysis (FHA) was used to predict the fertility of water buffalo bulls. FHA Model for buffalo was developed using the top 32% (conception rates = 28 to 36%) of the highest fertility bulls and the bottom 16% (conception rates = 19 to 13%) of bulls with the lowest fertility. The model cutoff points were harmonic amplitude 1 values of <0.042 µm and >0.051 µm for the high and low fertility bulls, respectively. Using the cutoff values, a new set of 44 bulls was classified and 12 high and 10 low fertility bulls identified. Five bulls from each fertility group were randomly selected and used to evaluate sperm in vitro fertility via fertilization of buffalo oocytes in vitro and sperm parameters via Computer Assisted Sperm Analysis (CASA) and Hypoosmotic Swelling Test (HOST). Buffalo oocytes were matured in vitro and fertilized with capacitated buffalo sperm for 18 hours from the selected bulls. Formation of a male pronucleus, cleavage, blastocysts development and hatching rate were used as evidence of fertilization and developmental competence of sired zygotes. There was a trend on HOST (68.0±6.2 vs. 59.1±6.2) and fertilization percentage difference (83±4.4 vs 70.0±4.4) for the predicted high vs. low fertility bulls. Significant difference (p<0.01) was observed in cleavage (73.5±0.5 vs. 60.1±0.7), blastocysts development (27.4±0.7 vs. 18.5±1.6), and hatching rates (38.3±1.2 vs. 28.7±2.7). The CASA measures of progressive motility (67.2±2.9 vs. 51.9±2.9), straightness (STR) (85.4±0.7 vs. 81.3±0.7) and LIN (53.3±0.8 vs. 49.8±0.8) were different for sperm from high vs low predicted fertility bulls (p<0.05). There was no effect of VAP, VSL, VCL ALH or BCF in relationship to bull fertility group (p>0.05). The results suggest that buffalo bulls predicted to be of higher fertility by FHA have better in vitro fertility and motion characteristics of progressive motility, STR and LIN. These results demonstrate efficiency of FHA as tool in predicting bull fertility.

The Philippine Department of Agriculture-Biotechnology Program Office provided the funding support of this research (DABiotech 1404). The Philippine Carabao Center and the University of Wisconsin-Madison provided the research facilities.

[1] Hufana-Duran D. 2009. Studies for the improvement of in vitro culture systems of oocytes and embryos in water buffaloes. http://www.tulips.tsukuba.ac.jp/limedio/dlam/B28/B2871346/1.pdf [2] Hufana-Duran D, EP Atabay, PG Duran, EC Atabay, Y Takahashi, Y Kanai, L. Cruz. 2008. Selection of water buffalo oocytes with developmental competence in vitro. Philippine Journal of Veterinary and Animal Sciences 34:149-158. [3] Hufana-Duran D, R Mallari, D Suba, P Duran, E Abella and F Mamuad. 2015. Hypo-Osmotic Swelling Test for Membrane Integrity Evaluation of Frozen-thawed Water Buffalo (Bubalus Bubalis, Linn.) Spermatozoa. Philippine Journal of Science 144(2):209-219. http://philjournalsci.dost.gov.ph/ [4] Jackson S, Brashers DE. Random Factors in ANOVA. Sage University Paper Series on Quantitative Applications in the Social Sciences, 07– 098. Thousand Oaks, Calif: Sage Publications; 1994: 1–65. [5] Ostermeier GC, Sargent GA, Yandell BS, Evenson DP, and Parrish JJ. 2001a. Measurement of sperm nuclear shape using Fourier Harmonic Analysis. J. Androl. 22:584-594. [6] Ostermeier GC, Sargent GA, Yandell BS, Evenson DP, and Parrish JJ. 2001b. Relationship of bull fertility to sperm nuclear shape. J. Androl. 22:595-603. [7] Parrish et al., 2014. Association for Applied Animal Andrology 2014 meeting. p30 – 49. In. International Veterinary Information Service, http://www.ivis.org/proceedings/aaaa/2014/3.pdf [8] Parrish J, Schindler J, Willenburg K, Enwall L, Kaya A. 2012. Quantitative sperm shape analysis: What can this tell us about male fertility. In: Proc. 24th Tech. Conf. Artif. Insemin. Reprod., National Association of Animal Breeders, Columbia MO. pp. 74-80.

43 ORAL ABSTRACTS Lalahan Model in Commercial Anatolian Buffalo Semen Production

Fırat KORKMAZ, Ilktan BAŞTAN, Derya ŞAHIN, Seher YIRTICI

International Center for Livestock Research and Training, Turkey [email protected], [email protected], [email protected], [email protected]

Due to their inheritable genetic heritage, regional compatibility, resistance, and immunity to local diseases, to protect and conserve our local breed Anatolian Buffalo is our national responsibility. In this context, commercial production of cryopreserved Anatolian Buffalo semen for using in artificial insemination and embryo transfer was started for the first time at International Center for Livestock Research and Training under the Directorate of Turkish Ministry of Agriculture and Forestry in 2017. Two Anatolian Buffalo bulls (3,4 years old) (Dülgeroglu and Karahisar) were selected to semen producing after various contagious diseases tests and quarantine under the control. The production began in regard to ‘Lalahan Model’ as we described a whole setup which includes semen collection with our self-made setup, semen packaging to 0.25 ml straws, freezing and storing final at -196 C in liquid nitrogen until the sale. It also comprises post-thaw subjective and objective semen quality analyses at 0h and 3h such abnormality, total (58,03±14,83, 3h=38,3±18,32) and progressive motility (24,20±8,22, 7,98±5,25), curvilinear velocity (VCL) (169,29±26,01, 125,7±29,04), average path velocity (VAP) (98,12±14,56, 69,93±15,73), straight-line velocity (VSL) (76,08±10,92, 50,51±7,99) and other sperm kinetic parameters by using computer-assisted sperm analyzer (CASA) and flow cytometric analyse (plasma membrane and acrosome integrity (PMAI (54,04±12,52, 38±11,83))) from each lot of products (0h=mean±std, 3h=mean±std). These results were collected and minimum limits were defined to eliminate low-grade lots before the sale. According to these results, 5000doses cryopreserved Anatolian Buffalo semen was produced with this mentioned ‘Lalahan model’ from 2017. In line with this model and gained experience from this preliminary study, our buffalo bulls stock is improving and we are continuing to the production of cryopreserved Anatolian Buffalo semen.

44 ORAL ABSTRACTS Retrospective of Ten Years of Fixed Timed Artificial Insemination (ftai) In Buffaloes in Amazon Region, Brazil: Preliminary Results

William VALE, Sebastião Tavares ROLIM-FILHO, Haroldo Francisco LOBATO RIBEIRO, Luís Américo BARBOSA, Henry Manrique DANIEL AYALA, Wilton LIMA, Loreno FRANCES, Anelise De Sarges Ramos RAMOS

Universidade Federal Rural da Amazonia, Brazil [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

The artificial insemination in buffaloes had its primordial in Latin America, in 1981, when deep frozen semen was locally processed and used successful for the first time, using natural estrus, when the first calves were born through this biotechnology (Vale et al. 1983). Since then, a lot of progress has been done and today FTAI is feasible biotechnology leaving a great legacy for the genetic improvement of buffalo in Brazil and other Latin American countries. The objective of this study was to evaluate the conception rates (CR) of female buffaloes submitted to (FTAI), with different synchronization protocols, during the last 10 years, in the Amazon Region, Brazil. A total of 2477 crossbred buffaloes (Murrah x Mediterranean) were submitted to FTAI, with the following synchronization adaptations, of which 1342 (54.1%) were pregnant, depending on the protocol used (table 1). The data were submitted to the chi-square test with 5% significance. The CR ranged from 38% to 58.5% (P <0.05). Protocols using progesterone presented a higher CR (58.5%) than protocols that do not use progesterone (38%). In addition, animals managed on cultivated pastures tend to have a better CR than animals inseminated in native pastures (52.9%). Therefore, factors such as type of protocol, use of progesterone and pasture type influence the CR of inseminated buffaloes through FTAI.

Table 1. Pregnancy rate of female buffaloes submitted to FTAI using different synchronization protocols, in Amazon Region, Brazil

Protocol N. Pregnant Canception rate (%) -Ovsynch: D0-GnRH; D7-PGF; D9-GnRH; D10-IATF 220 84 38,0% -*Progesterone com eCG - D0 (P4 + E2); D9 (eCG + PGF); D11 (GnRH); D12 (IA) 1250 731 58,5% -Progesterone sem eCG-D0 (P4 + E2); D9 (PGF); D11 (GnRH); D12 (IA) 818 427 52,2% -**Progesterone com eCG - D0 (P4 + E2); D9 (eCG + PGF); D11 (GnRH); D12 (IA) 189 100 52,9% Total 2477 1342 54,1%

*Extensive management in floodplain pasture, Marajó island. **Semi-intensive management in artificial pasture.

Literature Vale WG, Ribeiro HFL, Sousa JS, et al. Artificial insemination in buffaloes (Bubalus bubalis) in Amazon region. In: proceedings of the XXI Cong Bras Med Vet, Belém. 1984; 1:91.

45 ORAL ABSTRACTS Comparison of Growth Curve Models in Murrah Buffaloes

1Asli ARDIC, 2Serdar GENC, 1Erkan PEHLIVAN, 1Gursel DELLAL

1Ankara University, Agriculture Faculty, Animal Science Department, 06110, Ankara, Turkey 2*Kirsehir Ahi Evran University, Agriculture Faculty, Department of Agricultural Biotechnology, 40100, Kirsehir, Turkey *Corresponding Author E-mail: [email protected]

Dr. Öğr. Üyesi, Turkey [email protected]

In this study; Richards, Gompertz, Logistic, Von Bertalanffy and Brody Models was determined using by the relationship between age and body weight in male (n = 20) and female (n = 20) Murrah buffaloes raised in Turkey. The accuracy of estimations in the models were detected using by the R2 (coefficient of determination), R (corrected coefficient of determination) and MSE (mean squares errors). The five models used in both male and female were found a high hit rating in estimations of growing. However, the best estimate was obtained in Von Bertalanffy model (R2 = 99.88%) and the MSE in males and females were found as 134.26 and 123.71, respectively. Using this model, it was concluded that some growth and development characteristics such as mating age, breeding age and appropriate age of slaughter can be estimated in Murrah buffaloes.

46 ORAL ABSTRACTS Cooling Performance Assessment of Fogging System in Anatolian Water Fogging System with Regard to Animal Welfare

Mehmet Ihsan SOYSAL1, Israfil KOCAMAN2, Hüseyin Cömert KURÇ2

1Namik Kemal University Agricultural Faculty Animal Science, Turkey 2Namik Kemal University Agricultural Faculty Biosystems Engineering Department, Turkey [email protected], [email protected], [email protected]

This research was carried out to evaluate the cooling performance of fogging system which were installed in the water buffalo barn with regard to animal welfare. Two experimental groups which have ten water buffaloes for each group were formed based on genetic similarities, same number of lactations and same birthdates in the studied farm. The first group was housed in the Barn-I where the indoor temperature was controlled and the fogging system was installed, and the second group was housed in the Barn-II under existing farmer conditions. Same feeding ration was implemented for two groups. The cooling efficiency values of the fogging system installed in the Barn-I were varied between 45.6% and 85.7%. Temperature-Humidity Index values were 19.9 and 23.1 in the Barn-I and 19.5 to 26.4 in the Barn-II. As a result, a well-planned fogging system for animal welfare will be extremely useful in eliminating or reducing the negative effects of temperature stress.

47 ORAL ABSTRACTS Pregnancy-associated Glycoproteins and Early Pregnancy Failures in Buffalo Cows

Vittoria Lucia BARILE1, Laura MENCHETTI2, Gabriele BRECCHIA2, Claudio CANALI2, Olimpia BARBATO2

1CREA Research Centre for Animal Production and Aquaculture, Italy 2Department of Veterinary Medicine, University of Perugia, Italy [email protected], [email protected], [email protected], [email protected], [email protected]

It has long been accepted that pregnancy status should be determined in dairy cows as soon as possible so that non- pregnant animals can be reinseminate promptly to minimize the breeding economic losses. Pregnancy can be monitored using a variety of method including pregnancy-associated glycoproteins (PAG). (Fricke et al, 2016; Barbato and Barile, 2012). The plasma PAG profiles of water buffalo during pregnancy and postpartum periods have been recently described from our research group (Barbato et al., 2017; 2018). Aim of this work was to examine the relationship between PAG plasma concentrations and maintenance of pregnancy in early stage to evaluate embryo loss in buffaloes.

Ninety-five buffalo cows, belonging to a buffalo herd subjected to synchronization and artificial insemination (AI) program, were enrolled in this study. Blood samples were taken on days 0 (AI), 25, 28, and 45. Plasma was immediately separated by centrifugation and stored at -20 °C until assayed. PAG concentration was determined by radioimmunoassay systems (RIA-706) as reported previously by Barbato et al. (2018). Pregnancy was diagnosed by ultrasound on days 28 and 40 and confirmed by rectal palpation on day 60. The animals were classified ex post as pregnant (P), non-pregnant (NP) and that experienced embryo mortality within 28 and 40 days (EM). Only animals related to P and EM groups were included in the data analysis.

Thirty-four out of the 95 buffalo cows enrolled in this study became pregnant and maintained pregnancy (P group) while 12 have had embryonic mortality (EM group) and the others remained non-pregnant as determined by ultrasonography. After AI, concentrations of log-PAG-1 were higher in P group than EM group in each day of observation (mean difference: 0.3±0.1 ng/mL, 0.3±0.1 ng/mL, and 11.6±0.1 ng/mL at 25, 28, and 40 days post AI, respectively; P<0.01). Concentrations of PAG lower than 1.6 ng/mL, 2.2 ng/mL and 2.7 ng/mL at day 25, 28 and 40 allowed to discriminate between animals that maintained pregnancy (P group) and those that experienced embryo mortality (EM group).

[1] Barbato O, Barile VL. 2012. The pregnancy diagnosis in buffalo species: laboratory methods. J Buffalo Sci, 1 (2), 157- 162. [2] Barbato O, Menchetti L, Sousa NM, Malfatti A, Brecchia G, Canali C, Beckers JF, Barile VL. 2017. Pregnancy-associated glycoproteins (PAGs) concentrations in water buffaloes (Bubalus bubalis) during gestation and the postpartum period. Theriogenology, 97 :73-772. [3] Barbato O, Menchetti L, Sousa N, Brecchia G, Malfatti A, Canali C, Beckers JF, Barile VL. 2018. Correlation of two radioimmunoassay systems for measuring plasma pregnancy‐associated glycoproteins concentrations during early pregnancy and postpartum periods in water buffalo. Reprod Dom Anim, 53 (6): 1483-1490. [4] Fricke P M, Ricci A, Giordano J O, Carvalho P D. Methods for and Implementation of Pregnancy Diagnosis in Dairy Cows. 2016. Vet Clin North Am Food Anim Pract. 32:1, 165-180.

48 ORAL ABSTRACTS Investigation of Antimicrobial Susceptibility and Biofilm Forming Ability of Staphylococci from Subclinical Anatolian Buffalo Mastitis*

Ece KOLDAŞ ÜRER1, Özkan ASLANTAŞ2, Erhan TEK2, Mehmet Ali YILMAZ3, Yaşar ERGÜN4

1Hatay Mustafa Kemal University, Faculty of Veterinary Medicine, Department of Obstetrics and Gynecology, Antakya, Hatay, Turkey 2Hatay Mustafa Kemal University, Faculty of Veterinary Medicine, Department of Microbiology, Antakya, Hatay, Turkey 3International Center for Livestock Research and Training, Lalahan, Ankara, Turkey 41Hatay Mustafa Kemal University, Faculty of Veterinary Medicine, Turkey [email protected], [email protected], [email protected], [email protected], [email protected]

In this study, it was aimed (i) to determine the prevalence of subclinical mastitis in Anatolian buffaloes in Çorum province, (ii) to isolate and identify staphylococci involved, (iii) to determine the antibiotic susceptibilities of the isolates, (iv) to evaluate biofilm forming ability of the isolates by using Congo red agar (CRA) method and Microplate (MP) method, and (v) to investigate the genes responsible for biofilm forming ability and microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) by polymerase cahin reaction (PCR). To this end, 200 lactating buffalos were screened by California Mastitis Test (CMT), 48 (24%) were subclinical mastitis and 160 milk samples were collected for microbiological examination. Out of 160 milk samples, 107 (66.9%) Staphylococcus spp. were isolated, of which 26.2% were S. aureus and 73.8% were coagulase negative staphylococci (CNS) by MALDI-TOFF. The staphylococci were especially resistant to beta-lactams except cefoxitin, but were least resistant to other antibiotics tested. Based on CRA method, 92.9% of the S. aureus and 70.9% of the CoNS isolates were positive, whereas all S. aureus and 97.5% of CNS isolates were positive for biofilm forming ability by MP method. In S. aureus and CNS isolates, positivity rates for icaA and icaD genes were 14.3-1.3% and 64.3-19%, while the frequency of clfA, eno, clfB, ebpS, fib, fnbB, bpb and cna were 89.3-13.9%, 71.4- 75.9%, 71.4-12.7%, 67.9-10.1%, 64.3-7.6%, 28.6-7.6%, 3.6-0%, 0-10.1%, respectively. The bap gene was not detected in any staphylococci studied. The results indicated that staphylococci were mainly resistant to beta-lactams. In addition, it was detected that the presence of icaA and icaD genes was not always correlated with biofilm synthesis, and even in the absence of these genes, the isolates were able to synthesize biofilm. Therefore, there is a need not only for further studies for accurate evaluation of biofilm forming ability of staphylococci, but also for implementation of alternative methods for the effective treatment of infections caused by staphylococci.

*This study was supported by Hatay Mustafa Kemal University Scientific Research Fund (Project Number: 18 M 060)

49 ORAL ABSTRACTS Effect of Propolis In Water Buffaloes Cows with Subclinical Mastitis

Luıs MANTILLA, Raul CASANOVA, Rocio CASTRO

Laboratorio de investigaciones microbiológicas (LIM), Decanato de Investigación Universidad Nacional xperimental del Táchira (U, Venezuela) [email protected], [email protected], [email protected]

ABSTRACT: A study was carried out to prove the efficacy of treatments, based on propolis soft extracts, to control subclinical mastitis in water buffaloes cows in Parcela 7, located in the north region of Táchira state, Venezuela. The research had three phases: 1) Diagnosis to determine the presence of subclinical mastitis by performing California Mastitis Test (CMT) and microbiology test to detect etiological agents; 2) In vitro trials using Kirby-Bauer disk-susceptibility method; 3) in vivo trials by intramammary applications. The results showed an initial subclinical mastitis prevalence of 29.5%. Staphylococcus aureus was determined as the main etiological agent and therefore was isolated and used in the in vitro trials. The statistics means of the zone size of inhibition were: 21.33 mm for propolis at 50%; 20.11 mm for propolis at 25%; 24.44 mm for ciprofloxacin positive control; and 26.33 mm for the commercial product nitrofurantoin. These treatments were classified as “extremely sensitive” in the Duraffourd scale. For the in vivo trials the quarters diagnosed with subclinical mastitis were randomly distributed into three groups and received two repetitions for each treatment. The treatments used were: negative control; propolis at 50%; propolis at 25%; and commercial treatment. Propolis at 50% treatment healed 66.6% of the quarters and propolis at 25% healed 33.3% of the quarters which means that propolis treatments succeeded in 50% of the cases. The results allow to conclude that the solutions made of soft extract of propolis showed to be extremely sensitive in the in vitro trials and were effective in the control of the subclinical mastitis caused by staphylococcus aureus in water buffaloes cows of the north region of Táchira state.

[1] Bauer AW, Kirby WM, Sherris JC, Turck M.1966. Antibiotic susceptibility testing by a standardized single disc method. American Journal of Clinical Pathology, 45: 493. [2] Bačić G, Macesic N, Radin, L, Aladrovic, J, Matanovic K. 2016. Intramammary Propolis Formulation for Subclinical Mastitis Prevention and Treatment in Dairy Cows. J Dairy Vet Anim Res 3(5): 00091. DOI: 10.15406/jdvar.2016.03.00091. [3] Bogdanov S. 2017. Propolis: Composition, Health, Medicine: A Review. [4] Duraffourd C, D`hervocourt L, Lapraz JC. 1986. Cuadernos de Fitoterapia Clínica. 1ª edición. Barcelona, España:Edit. [5] Neacato S (2005). Uso de extractos etanólicos de propóleos para el control de Staphylococcus aureus “in vitro” obtenido de leche de vacas con mastitis. Trabajo de grado no publicado. Escuela Politécnica de Barquisimeto. Ecuador. [6] Kushikawa E, Mello E, Maiklei R, Sussumu L. 2011. mastitis bovina por Staphylococcus aureus: sensibilidade as drogas antimicrobianas e ao extrato alcoolico de propolis [7] Najmadden H, kakamand F. 2009. Antimicrobial activity of propolis collected in different regions of Sulaimani province- Kurdistan region/Iraq j Duhok univ.12(1):233-239. [8] 8. Fiordalisi S, Honorato L, Loiko M, Avancini C, Veleirinho M, Machado L, Kuhnen S. 2015. The effects of Brazilian propolis on etiological agents of mastitis and the viability of bovine mammary gland explants. Journal of Dairy Science, Vol. 99, Issue 3, p2308–2318 [9] Freitas Santana H., Teixeira Barbosa A, Olavo Ferreira S, Mantovani H. 2012. Bactericidal activity of ethanolic extracts of propolis against Staphylococcus aureus isolated from mastitic cows. [10] Zambrano M. 2012. Guía de prácticas de laboratorio. Microbiología aplicada.

50 ORAL ABSTRACTS Water Pumps Moved by Draught Animal Power Versus Water Pumps Moved by Fuel Engines or Photovoltaic: A Cost Benefit Analysis

Antonio PERRONE

WEDAP, Italy [email protected]

In the rural areas hundreds of thousands of pumps, threshers, sieves, mills and milking machines are moved by hand or pedal cranks. In the same areas hundreds of millions animals are working in agriculture for land cultivation and transports. Relevant data on the matter are available in Integrated Crop Management - Mechanization for Rural Development: A review of patterns and progress from around the world (Vol. 20-2013), a FAO publication. Several scientific institutions, mainly in Asia, are engaged in the development of motors, moved by draught animal power, to supply energy to the said equipment now moved by hand or pedal cranks. The way taken has been a system in which the animal, walking in a round path, moves a rounds multiplier that moves a generator of electric current.

The electricity produced then moves pumps, sieves, threshers and other rural equipment. It is well known that the average muscular energy of a working man is of one hundred watts while the average muscular energy of a medium sized draught animal is of five hundreds watts. This is evident in land cultivation: a farmer who hoes the land is five times less efficient than a farmer that plows the land with the support of a buffalo or an ox. The same increase of productivity can be now reached for the stand alone plants such as pumps and the other equipment above mentioned. The new electric systems have the following advantages with respect of the traditional mechanical systems: an easier transmission of the energy form the animal to the pumps (by cable and not through gears) or the other equipment, an easier supply of the equipment that are the same utilized with fuel generators or with the connection to the electrical grid. A feasibility study has demonstrated, through the calculation of the net present value” that the cost of the use of draught animal power is cheaper than the cost of the manual labor, of the fuel engines, of the photovoltaic, of the connection to the electrical grid and of the wind generators.

Thanks to an agreement, between the Mechanization Unit of the Ente CRA, the Research Council of the Italian Ministry of Agriculture and the International Buffalo Federation, one of the prototypes realized by Antonio Perrone has been tested in Monterotondo, Rome. The Unit has issued a report on the test run the July 14th , 2011.

[1] Antonio Perrone has been for several years the head of division of the Directorate for Sustainable Development of the Ministry of the Environment in Italy. [2] Thanks to a scholarship of the Italian National Council of Researches, he has been Visiting Scholar at the New York University School of Law. He has held two Masters, in science and technology management, at the Governmental School of Public Administration in Italy. [3] The Water Division of FAO has hosted Antonio Perrone, in Sirte in Libya in December 2008, on the occasion of the Ministerial Conference on Water and Energy for Agriculture in Africa. [4] CETAMB, “CeTAmb LAB – Laboratorio di ricerca sulle tecnologie appropriate per la gestione dell’ambiente nei Paesi a risorse limitate”, a center of Brescia University directed by prof. Carlo Collivignarelli, on June18th, 2014 organized a Workshop The Draught Animal Power: a tool for development not so developed to discuss the work done till now by Antonio Perrone within the initiative here presented. [5] Antonio Perrone presented a speach on June19th, 2014 at the 15 days IBF TRAINING COURSE ON BUFFALO MANAGEMENT AND INDUSTRY organized by the International Buffalo Federation in Latina and attended by 27 professional from 12 countries of 3 continents. [6] The Asian Buffalo Congress 2015 (Istambul 21-25 April 2015) hosted his poster THE DRAUGHT ANIMAL POWER: A DEVELOPMENT TOOL NO SO DEVELOPED

51 ORAL ABSTRACTS Introduction of Nikookar Buffalo Farm

Fatemeh DERISAVI

Cooperative of Bufalo Breeders, United States [email protected]

Nikookar buffalo farm is located in Rezvanshahr city,400 km far from Tehran, Gilan province(North west of Iran and south of Caspian sea). Because of my interest in buffalo breeding, along with my husband, we did a reverse migration from city to village in 1999. In the beginning we started to work with ten local buffaloes in traditional system. After several years our buffaloes increased but we understood that traditional system was not economic and profitable. Therefore, we decided to improve our herd through new methods of breeding mainly to use artificial insemination of Italian frozen semen, along with embryo transfer of Italian frozen embryo and by these methods we could improve our herd. At present, I have fifty five (55) heads of F1 , F2 and F3 crossbred buffaloes which all of them are milked by machine and calves are milk fed by artificial teat. But still I was not satisfied and decided to serve and train other small size farmers of my country. In this connection, I transferred to them all of my experiences, in order to achieve the best result, I established the first successful cooperative of buffalo breeders in Iran in 2017.So that, at present our cooperative has 150 (Hundred fifty) members. In connection with these new methods among other small size farmers I used the following ways:

1-To train on the establishment of cheap buffalo housing with regarding technical and sanitary standards. 2-To train milking buffaloes by machine through the field visit of farmers from my own farm. 3-To train the preparation of balanced ration with emphasis on usage of wastes and residues with the least cost. 4-To transfer of superior genes to other farms through selling of my own F1, F2 and F3 animals to them. 5-To train the correct and hygienic ways of milking in order to decrease total count, thereby, better market for their milk. 6-To distribute cheap inputs among members of our cooperative. 7-To collect and guarantee purchase of milk by cooperative in order to increase their income. 8-To train processing of rural women in order to better sale and better welfare so that the cheese produced by myself in my farm has many customers in all over the country.

In the end, I believe to establish the pilot buffalo farm in rural areas of countries similar to Iran has significant effect in improving of farms and sustainable development in rural communities. Thereby, this animal of third millennium based on FAO statement can create suitable food security for mankind.

52 ORAL ABSTRACTS Sustainable Buffalo Production: A Case Study from Nepal

Nanda JOSHI, Ramjee GHIMIRE

Department of Animal Science, Michigan State University, United States [email protected], [email protected]

Buffalo population of Nepal is estimated at 5 million ~ 2.6% of world buffalo population (FAOSAT 2019). Due to its ability to utilize low quality forages and pastures, relatively resistant to diseases, and adaptable to a wide range of housing, feeding, and management conditions (Wanapat and Kang 2013) it is popular specie amongst the Nepalese smallholder farmers. Nepalese famers, on average, keep 2-5 buffaloes on less than one ha of land. It contributes more than 70% of milk and 65% of meat production that is about 50% of livestock GDP (Hada 2014, Nirmal 2017). Winter fodder scarcity is the major problem in feeding management for ruminants (Devkota et al. 2015). Forage scarcity is aggravated by the fact that most pasturelands are overused and undermanaged. Increasing effect of climate change include; prolonged drought, uneven distribution of monsoon rain that causes decline in forage and pasture productivity impacting buffalo production and productivity (Joshi 2018). Grounded on case studies from Nepal, this paper discusses the lessons learnt from projects on smallholder buffalo production systems, including effects of climate change on feed and forage availability; breeding, feeding, pasture/rangeland management; housing management; livestock and human nexus and zoonoses (Kelly et al. 2018); and social and economic issues (Muralikrishna and Manickam, 2017) on buffalo production. This paper also highlights the interconnectivity of these issues and the need to examine and address issues in a holistic manner.

A road map for sustainable buffalo development in Nepal is discussed that include; emphasis on policies to discourage slaughtering of female buffaloes after one or two calving’s; improve meat quality and hygienic conditions; and improve housing. It is strongly felt that any efforts to develop buffalo sector should be participatory, informing and empowering buffalo farmers so that they become active partners in the process, own the outcomes, and identify and pursue appropriate development strategies in the future.

[1] FAOSTAT. 2019. Data. http://www.fao.org/faostat/en/#data [2] Hada, N. 2014. Cattle and Buffalo: Emerging Opportunity. USAID Nepal. [3] Joshi, N. 2018. Adaptation of buffalo production system to the impact of climate change: A case study from Nepal. [Abstract]. In S. K. Khurana et al. (Eds). Book of Abstracts: ABC -2018. [4] Kelly, T. R., Bunn, D. A., Joshi, N., Grooms, D., Devkota, D., Devkota, N. R., and Mazet, J. A. 2018. Awareness and Practices Relating to Zoonotic Diseases Among Smallholder Farmers in Nepal. EcoHealth, 15(3) 656-669. [5] Nirmal, B. K. 2017. Strategy to enhance the production and productivity of buffaloes for the growth of Livestock Gross Domestic Product (LGDP) of Nepal. Kathmandu, Nepal.

53 ORAL ABSTRACTS Description and Typology of Water Buffalo Enterprises in Marmara Region, Turkey

Nurşen ÖZTÜRK1, Ferhan KAYGISIZ1, Pembe Dilara AKIN1, Hülya YALÇINTAN1, Mehmet Ihsan SOYSAL2, Mustafa KÜÇÜKKEBABÇI3, Ömür KOÇAK1

1Istanbul University-Cerrahpasa, Turkey 2Tekirdag Namik Kemal University, Turkey 3Sheep Breeding Research Institute, Turkey [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

Water buffalo farming in Turkey generally is conducted by low yield animals with traditional rearing methods and small scale-family farms are the dominant type of these rearing systems. However, in the last decade as a result of rapid urbanization, consumers’ consciousness has risen on dietary topics and animal-origin products has begun to be demanded more. Mozzarella and ricotta cheese, yoghourt and milk cream (kaymak) are the most demanded products made by buffalo milk. The increase in consumer demand and consumers’ willingness to pay for higher prises to buffalo products created a change in production systems. In order to increase the amount of milk sold some farmers has started to rear Italian buffaloes instead of Anatolian buffaloes and started to apply intensive rearing systems, such as using more concentrate. This study aims to investigate the different production systems in water buffalo rearing in Marmara Region of Turkey. We plan to conduct a survey study in Istanbul, Kocaeli, Balikesir and Kirklareli cities with fifty buffalo farm owners in order to identify the variation within the buffalo rearing. For this purpose, we will investigate the land use, herd structure, buffalo feeding, animal health, production and economic performance, labour use, investment in buildings and equipment, and farmer demographic parameters and identify the variation exists in the buffalo rearing. We will perform Principal Component Analysis to identify differences among farmers and later we will apply Cluster Analysis to identify farm groups in Marmara region. Finally we will identify the technical and economic differences among farm clusters by applying ANOVA and Chi-Square analysis.

The study will demonstrate the strong and weak points of the farm clusters and outcomes of this study will directly support the policy makers in terms of their livestock supporting policies. We would like to thank to Republic of Turkey Ministry of Agriculture and Forestry General Directorate of Agricultural Research and Policies, Şenol Çakır, Talat Tan, Mehmet Aksel and Rabiya Birkiye.

54 POSTER PRESENTATIONS Crea Research Activity on Biogas Production from Buffalo Wastes

Antonella CHIARIOTTI1, Serafino CONCETTI1, Alessandra CRISÀ2

1CREA Research Center for Animal Production and Aquaculture, via Salaria 31, 00015 Monterotondo, Italy 2CREA, Italy [email protected], [email protected], [email protected]

Anaerobic digestion of agricultural and industrial wastes delivers two benefits: on one side, the treatment of organic residues prevents the environmental and economic impact of their disposal; on the other side, methane and/or hydrogen are generated from renewable sources. Current interest in the use of hydrogen as fuel, both for industrial applications and for road haulage, is because hydrogen-produced pollution is almost nil. When used in combustion systems, hydrogen only produces water and traces of N oxides; while if it is used with electrochemical systems (fuel cells) it produces only water. Various factors influence H2 production such as substrate, microbial inoculum, pretreatment, pH, temperature.

The CREA, Research Center for Animal Production and Aquaculture has been involved in anaerobic digestion and dark fermentation studies for few years now. Laboratory trials and microbiological studies have been undertaken, as well as the upgrading of a two-stage pilot plant for combined production of methane and hydrogen working at mesophilic conditions. The advantages of this technology are: animal waste recycling to produce energy (H2, CH4); higher methane production; additional hydrogen production; avoid food-no food competition; GHG reduction; smaller size biogas plants.

The lab trials carried out at CREA using buffalo wastes (sludge and cheese-factory by-products) are presented and discussed. This work was supported by the Italian Ministry of Agriculture (MiPAAF) under the AGROENER project (D.D. n. 26329, 1 april 2016) - http://agroener.crea.gov.it/

55 POSTER PRESENTATIONS Welfare Assessment of Buffalo Cows: Use of Expert Opinion Elicitation for The Characterization of Hazards, Welfare Promoters and Animal-Based Measures

Domenico VECCHIO1, Pasquale ROSSI2, Carlo GRASSI2, Pasquale RUSSO2, Lazzaro IEMMA2, Domenico CORONATI2, Massimo PALLADIO2, Franco CONSALVO3, Giuseppe DE ROSA4, Fabio NAPOLITANO5, Gianluca NEGLIA6, Esterina DE CARLO7, Valentina LORENZI8, Francesca FUSI8, Luca BOLZONI9, Stefano PONGOLINI9, Giorgio GALIERO10, Giovanni Loris ALBORALI11, Luigi BERTOCCHI8

1National Reference Centre on Water Buffalo Farming and Productions Hygiene and Technologies IZSM, Portici,, Italy 2Freelancer Veterinarian, Italy 3Veterinarian ASL Salerno, Italy, Italy 4Dipartimento di Agraria, Università degli Studi di Napoli Federico II, Portici, Naples, Italy 55 Scuola di Scienze Agrarie, Forestali, Alimentari ed Ambientali, Università degli Studi della Basilicata, Potenza, Italy 6Universty of Naples Department of Veterinary Medicine and Animal Production, Italy 7National Reference Centre on Water Buffalo Farming and Productions Hygiene and Technologies IZSM, Portici, Italy 8Centro di Referenza Nazionale per il Benessere animale Istituto Zooprofilattico Sperimentale della Lombardia ed Emilia Romagna, Italy 9Unità Analisi del Rischio ed Epidemiologia Genomica, IZSLER, Parma, Italy, Italy 10Istituto Zooprofilattico Sperimentale del Mezzogiorno, Portici, Italy 11Centro di referenza nazionale per il benessere animale Istituto Zooprofilattico Sperimentale della Lombardia ed Emilia Romagna, Italy [email protected], [email protected], [email protected], [email protected], lazzaro.iemma82@ gmail.com, [email protected], [email protected], [email protected], [email protected], fabio. [email protected], [email protected], [email protected], [email protected], francesca.fusi@ izsler.it, [email protected], [email protected], [email protected], [email protected], [email protected]

An expert opinion elicitation, based on a modified Delphi technique, was organized to collect the opinion of 14 Italian veterinarians with the aim of conducting a hazard and a welfare promoter characterization for defining and weighing a list of management and housing factors potentially associated with negative or positive welfare outcomes in buffalo kept in loose housing systems. In addition, the 14 experts judged a set of animal-based measures (ABMs), rating their appropriateness and the level of animal pain and suffering due to the welfare consequences they measure. Experts were asked to score 52 hazards, 45 welfare promoters and 14 ABMs.

Management and housing hazards, that were determined to be associated with a very high negative impact on the welfare of buffalo, were: lack of water availability, uncorrected animal grouping strategy, dirty and unmanaged lying area, incorrect milking routine, poor maintenance of the milking system, lack of litter for calves, insufficient shaded lying area and/or lack of any cooling system and presence of inadequate or slippery floor in walking areas. Management and housing welfare promoters dealing with optimal buffalo comfort around resting obtained the highest ratings, in particular: housing free for all animals and the possibility for adult buffaloes to access a loafing area of at least 7 m²and/or to a pasture for at least 60 days a year; cleanleness of the calf boxes, a hospital pen of at least 8 m²/parturient animal. In addition, high scores were given to appropriate grouping, adequate temperature, humidity and ventilation, presence of experienced and trained stockpersons. As to ABMs, the highest rankings were given to mortality of calves and adult buffaloes, percentage of animals showing vaginal and/or uterine prolapses, body condition scoring and percentage of deformed claws in adult buffaloes. Our results represent the starting point for the development of the first protocol for the assessment of the welfare of buffalo farmed in loose housing systems.

Acknowledgements: This research was supported by Italian Ministry of Health, (number project cup: E52I14001190001)

[1] Bertocchi, L., Fusi, F., Angelucci, A., Bolzoni, L., Pongolini, S., Strano, R.M., Ginestreti, J., Riuzzi, G., Moroni, P., Lorenzi, V., 2018. Characterization of hazards, welfare promoters and animal-based measures for the welfare assessment of dairy cows: Elicitation of expert opinion. Prev. Vet. Med. https://doi.org/10.1016/j.prevetmed.2017.11.023 [2] De Rosa G., Grasso F., Winckler C., Bilancione A., Pacelli C., Masucci F., Napolitano F., 2015. Application of the Welfare Quality protocol to dairy buffalo farms: Prevalence and reliability of selected measures. J Dairy Sci. 2015 Oct; 98(10):6886-96. doi: 10.3168/ jds.2015-9350. Epub 2015 Jul 29. [3] EFSA Panel on Animal Health and Welfare (AHAW); Guidance on risk assessment for animal welfare. EFSA Journal 2012; 10(1):2513. [4] Hultgren, J., Algers, B., Atkinson, S., Ellingsen, K., Eriksson, S., Hreinsson, K., Nordensten, L., Valtari, H., Mejdell, C.M., 2016. Risk assessment of sheep welfare at small-scale slaughter in Nordic countries, comparing with large-scale slaughter. Acta Vet. Scand. 58, 34. [5] Ribó, O., Serratosa, J., 2009. History and procedural aspects of the animal welfare risk assessment at EFSA. In: In: Smulders, F.J.M., Algers, B. (Eds.), Welfare of Production Animals: Assessment and Management of Risks–Food Safety Assurance and Veterinary Public Health, vol. 5. Wageningen Academic Publishers, the Netherlands, pp. 305–335. [6] Smulders, F.J.M., 2009. A practicable approach to assessing risks for animal welfare – methodological considerations. In: In: Smulders, F.J.M., Algers, B. (Eds.), Welfare of Production Animals: Assessment and Management of Risks–Food Safety Assurance and Veterinary Public Health, vol. 5. Wageningen Academic Publishers, the Netherlands, pp. 239–274. [7] Vecchio D; Tamaio J.E., Bertocchi L; De Carlo E. 2017. Development of a new approach to evaluate Buffalo Welfare, the Ruminant Welfare® project. XVII Latin American Congress Buiatrics, Medellin, Colombia from 1-3rd November 2017. Invited lecture

56 POSTER PRESENTATIONS Are the Tomato Pomace Residues Useful for Hydrogen Production?

Antonella CHIARIOTTI1, Emanuela ROSSI1, Enrico SANTANGELO2

1CREA Research Center for Animal Production and Aquaculture, via Salaria 31, 00015 Monterotondo, Italy 2CREA Research Centre for Engineering and Agro-Food Processing Via della Pascolare, 16,00015 Monterotondo, Italy [email protected], [email protected], [email protected]

Tomato pomace is the main solid waste stream from tomato industry, and comprises peels, seeds and pulp (in a percentage depending on the specific process) which are separated before and during processing. In Italy around 3 to 6 Mt of residual biomass from tomato industry are produced each year (Calabrò, 2015). Such by-products were proved suitable for anaerobic digestion (Schievano et al., 2009) where their use in codigestion would be a rational alternative to the disposal. Limited research has been conducted to investigate the potential of using tomato pomace to produce hydrogen alone or in codigestion with buffalo sludge. Two batch experiments in an Automatic Methane Potential Test System (AMPTS, Bioprocess Control, Sweden) were conducted to verify the effect of pH pretreatment (adjusted and not adjusted at pH 9.0) on hydrogen production of tomato pomace (peels plus seeds) codigested with buffalo sludge at different ratio (1:2, 1:3, 1:4) of volatile solids (VS) concentration.

The hydrogen production was very scarce (5.36 % max) and only when the tomato pomace:buffalo sludge VS ratio was at 1:2. No production was observed with tomato pomace alone due to acidification (pH<3.5) of the batch. The pH pretreatment of tomato pomace did not affect biogas production. Total volatile fatty acids (VFA) concentration at the end of the process did not reveal any statistical difference. Nevertheless, the value of the 1:2 ratio was the highest (130, 127, 105 mM for 1:2, 1:3, 1:4 ratio respectively, P>0.05); acetate, propionate, and butyrate showed the same tendency. Small amounts of lactate were observed only with tomato pomace alone. Methane production was observed only in buffalo sludge batch because the pH drop caused by the peels was detrimental.Tomato pomace may have a great potential to be recycled rather than disposed. From this first approach the suitable VS concentration of tomato pomace appears the key factor to be careful determined for their exploitation when codigested with buffalo sludge.

This work was supported by the Italian Ministry of Agriculture (MiPAAF) under the AGROENER project (D.D. n. 26329, 1 april 2016) - http://agroener.crea.gov.it/

[1] Calabrò, P.S., Greco, R., Evangelou, A., & Komilis, D.P. (2015). Anaerobic digestion of tomato processing waste: Effect of alkaline pretreatment. Journal of environmental management, 163, 49-52. [2] Schievano, A., D’Imporzano, G., & Adani, F. (2009). Substituting energy crops with organic wastes and agro-industrial residues for biogas production. Journal of environmental management, 90(8), 2537-2541.https://doi.org/10.1016/j. jenvman.2009.01.013

57 POSTER PRESENTATIONS Method to Measuring Methane Emission in Italian Buffalo

Emanuela PARLATO1, Chiara CASO1, Angelo COLETTA2

1RISBUFALA, Italy 2risbufala, Italy [email protected], [email protected], [email protected]

Methane is a greenhouse gas with the global warming potential 25 times that CO2 (Forster et al., 2007). Therefore methane accounts for a great part of the emitted CO2-equivalents from agriculture. In ruminant animals, typically the most of the methane (>90%) is generated in the rumen during microbial fermentation of cellulosic feed materials (enteric methane) and a small part by a composition of manure (Verge et al., 2007). Globally the dairy sector is estimated to contribute 4% of greenhouse gas emission (FAO, 2010). Methane accounts for up to 50% of greenhouse gas emission from milk production, and its contribution can be up to 80% in in grassland systems (FAO, 2010). The total methane emissions on farm or on national level are calculated as the product of number of buffalo and emissions per buffalo. Therefore, develop a technique to accurately measuring methane emission in buffalo on farm, is of extreme importance. Several direct and indirect methods have been developed in the last years to measuring and estimating methane emissions in ruminants. They can have different applications, advantages and disadvantages. One of the most suitable techniques to measure methane emissions in ruminants under commercial conditions was reported by Garnsworthy et al. in 2012. This technique involves measurement of methane concentrations in air expelled through eructation by individual cow during milking. A designed programme is used to identify and quantify peaks. Raw data from the logger are transformed into values for peaks height and integral area under the peaks to measuring the total individual methane emission at eructation. This innovative technique will provide the opportunity to measuring methane emissions in dairy buffalo in accurate and economic way. And to using those data for calculation of genetic index for methane emission that will allow buffalo farmers to selecting for low emissions buffalo

[1] Forster, P., et al. Changes in atmospheric constituents and in radiative. Fourth Assessment Report of the Intergovernmental Panel on Climate Change. [2] H.L. Miller (Eds.) Climate Change. 2007: The Physical Science Basis. Cambridge University Press, Cambridge, UK and New York, NY; 2007:129–234. [3] FAO. FAOSTAT, 2018. [4] Goopy, C J.P.. Chang and N. Tomkins. A Comparison of Methodologies for Measuring Methane Emissions from Ruminants. Chapter 5. Standard Assessment of agricultural mitigation potential and livelihoods. 2016. Page 1-21. [5] Garnsworthy P.C., et al. On farm methane measurements during milking correlate with total methane production by individual dairy cows. 2012. J. Dairy Sci. 95:3166-3180. [6] Grainger, C., et al. Methane emissions from dairy cows measured using the sulfur hexafluoride (SF6) tracer and chamber techniques. 2007. J. Dairy Sci. 90:2755–2766. [7] RISBUFALA . www.risbufala.org. 2019. [8] Verge, X.P.C., Dyer, J.A., Desjardins, R.L., Worth, D. Greenhouse gas emissions from the Canadian dairy industry, 2001. 2007. Agric. Syst. 94: 683–693.

58 POSTER PRESENTATIONS Application with A Practical Selection Method of Buffalo Calves

Yusuf KAPLAN1, Samet CİNKAYA2, Mustafa DEMİRTAŞ2, Mustafa TEKERLİ2

1General Directorate of Agricultural Research and Policies, Ministry of Agriculture and Forestry, Turkey 2Afyon Kocatepe University, Faculty of Veterinary Medicine, Department of Animal Husbandry, Turkey [email protected], [email protected], [email protected], [email protected]

The selection of animals carrying higher breeding values is economically very important in the aspects of production and reproduction. Some complex models and methods are used for estimating the real breeding values in animal husbandry. But the conventional buffalo breeders need to some easy to use procedures for evaluating their dairy and meat animals. The aim of the study to demonstrate a practical method with a simple application. A computer software called Damızlık Asistanı (Assistant for studs) developed to National Anatolian Buffalo Breeding Project in farm condition were used for this purpose. Adjusted data by the software were treated according to a method named easy index and the results were used for selection of animals in the project (TAGEM/66MANDA2015-01) applied with the coordination of General Directorate of Agricultural Research and Policies, Republic of Turkey Ministry of Agriculture and Forestry in the province of Yozgat.

The author thanks to The General Directorate of Agricultural Research, Republic of Turkey Ministry of Agriculture and Forestry and Policies because of their support for the National Anatolian Buffalo Breeding Project.

59 POSTER PRESENTATIONS Effects of Year and Location on Lactation Milk Yield in Anatolian Water Buffalo In Giresun Province

Mustafa UĞURLU1, Ismail KAYA2

1Department of Animal Breeding and Husbandry, University of Ondokuz Mayis, Veterinary Medicine Faculty, Samsun, Turkey 2Depatment of Animal Nutrition and Nutritional Diseases, University of Ondokuz Mayis, Veterinary Medicine Faculty, Samsun, Turkey [email protected], [email protected]

This research was conducted to determine the effects of year and location on lactation milk yield in Anatolian Water Buffalo. The data on lactation milk records, belong to same buffalos, were taken from between 2013-2015 records of the Anatolian Water Buffalo Breeding Association in Giresun Province in Turkey. The effects year and location on lactation milk yield were assessed. The records, year was divided into three groups; 2013, 2014 and 2015. Location was divided into two groups; Dereli and Sebinkarahisar. Least square analyses were used for statistical analyzes. Statistical significance among subgroups were determined with the Duncan test. The overall means of total milk yield was 939.21 kg. In this study, lactation milk yield was significantly influenced by year (P<0.05) and location (P<0.05), and it was tend to increased with the year. In terms of location, the lactation milk yields in Sebinkarahisar (987.69 kg) were higher than those of Dereli (890.74 kg). In conclusion, lactation milk yield in Giresun province is satisfactory for Turkish conditions. In terms of year, increasing of lactation milk yield can be explained with increasing age of buffalo. However, location effect on lactation milk yield should be investigated in terms of management and climatic conditions.

This research was supported by Republic of Turkey Ministry of Agriculture and Forestry General Directorate of Agricultural Research and Policies

60 POSTER PRESENTATIONS Some Yield Features of Anatolian Water Buffaloes Grown in Diyarbakır Province by Local Stock Breeders

Yavuz HAN1, Mehmet Emin VURAL1, Ahmet KARATAŞ1, Bırusk KESKİN1, Rıdvan YAKİŞAN1, Galip BAKIR2

1Gap International Agricultural Research And Training Center, Turkey 2Sütçü İmam University, Turkey [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

This study was conducted in Anatolian Water Buffalo herds grown in Diyarbakir province. Breeding of Anatolian Water Buffalo was started in 21 villages with Anatolian Water Buffalo in 2011. The project has been started with 80 individual Anatolian Water Buffaloes of 2 years and older. Parameters that have been evaluated during the breeding project was milk yield, birth weight and body weight gain for the breeding materials. The mean birth weight of calves of Anatolian Water Buffalo was 30,01±0,15 kg in males (632 heads) and 29,73±0,14 kg in famales (709 heads) in the region, in 2017. Average of 6th month weight of calves were 91,82±0,78 kg in males (476 heads) and 90,79±0,64 kg in females (576 heads), in 2017. Average of 12th month weight of calves were 140,46±1,24 kg in males (317 heads) and 138,3±0,98 kg in females (440 heads), in 2017. The average lactation milk yield of Anatolian Water Buffaloes was 810,05±10,29 kg and the average lactation period of Anatolian Water Buffaloes was 185,59±1,31 days, in 1254 heads, in 2017.

61 POSTER PRESENTATIONS Investigation of Milk Yield by Using Relationships Between Body Measurements and Milk Yield In Anatolian Buffaloes With Multiple Regression

Serdar GENÇ1, Mehmet AKSEL2, Mehmet Ihsan SOYSAL3, Eser Kemal GURCAN3

1Kirsehir Ahi Evran University , Turkey 2Istanbul Anatolian Buffaloes Breeding Association, Turkey 3Namik Kemal University Agricultural Faculty Animal Science, Turkey [email protected], [email protected], [email protected], [email protected]

In this study, the relationships between body measurements and milk yield in Anatolian buffaloes were determined by regression analysis. For this purpose, measurements taken from a total of 45 Anatolian buffaloes, which are grown in the districts of Istanbul and included in the scope of Anatolian Buffaloes Breeding Project in Istanbul were used. In this study, withers height (WH), body depth (BD), body length (BL), rump width (RW), haunch lump width (HLW), chest width (CW), front nipple length (FNL), rear nipple length (RNL), front nipple connection (FNC), rear nipple connection (RNC), front nipple height (FNH), rear nipple height (RNH), nipple height (NH), rear leg rear view (RLRV), rear leg side view (RLSV) were among the body measurements and first lactation lenght (FLL) and first lactation milk yield (FLMY) were among milk yield properties were emphasized. Multiple regression analysis was used to estimate FLMY by using body measurements in Anatolian Buffaloes. In multiple regression analysis, it was tried to find the best model by using Mallows Cp statistic (MC) and accuracy (R2), while independent variables that could be included in the model according to the best subset regression method were found. According to the results of the regression analysis, the dependent variable is FLMY and 16 other variables (WH, BD, BL, RW, HLW, CW, FNL, RNL, FNC, RNC, FNH, RNH, NH, RLRV, RLSV, FLL) as independent variables has been added model. As a result, when the dependent variable FLMY and 16 other variables were added to the model independently, R2 = 0.478 and MC = 17. When these statistics are evaluated, it is difficult to estimate FLMY with body measurements by this method. Therefore, various genetic/molecular genetic or breeding methods can be used because of good estimation and more reliable results.

62 POSTER PRESENTATIONS Isolation of Streptococcus Uberis In Water Buffaloes Aborted Fetuses.

Luisa D’ANGELO, Domenico ALFANO, Roberto SCOGNAMIGLIO, Estrerina DE CARLO, Domenico VECCHIO

National Reference Centre on Water Buffalo Farming and Productions Hygiene and Technologies Istituto Zooprofilattico Sperimenta, Italy [email protected], [email protected], [email protected], [email protected], [email protected]

This work reports cases of isolation of Streptococcus uberis (Su) strains in three buffaloes fetuses aborted in Campania farms, southern Italy. This study aims to characterize the Su strains and to elucidate the antibiotic susceptibility profiles of the strains.

The isolation occurs following autopsy of aborted fetuses with culture test, using direct seeding of diagnostic swabs on general and notselective differential media (Blood agar 5%, Macconkey agar). Each fetus is also subjected to biomolecular analysis (isolation in PCR for: Leptospirosis, Bohv-1 and Buhv-1, Chlamydophila spp, Neospora caninum, Toxoplasma gondii, Coxiella burnetii ).

One fetus also results positive for Coxiella burnetii (isolation in RTpcr), another to Bacillus licheniformis and only in a fetus Su be isolated without other positivity. In first one, the anatomopathological examination shows vascular-type lesions with serum hemorrhagic effusion in the abdomen and thorax. The second one phenomena of autolysis against the parenchymal organs. The fetus with only Su shows no obvious pathological lesions. All three isolates of S. uberis have resistance to several molecules of antibiotics: nalidixic acid, Apramycin, Colistin sulfate, Flumequina, Kanamycin, Neomycin, Oxitetracycline, Tetracycline. The common sensitivity is only for molecules of Cephalissin, Florfenicol.

Although S. uberis is found as an opportunistic pathogen, it is also recognized as a potential primary and secondary abortion agent.

[1] P.J.Quinn, B.K.Markey M.E.Carter, W.J.Donnelly and F.C.Leonard, Veterinary Microbiology and Microbial Disease 2002. [2] E. W. Koneman- Testo Atlante di Microbiologia Diagnostica-1995 sec. Edizione. [3] D. Scott McVey, M. Kennedy, M.M. Chengappa – Veterinary Microbiology (2013), Ed. Wiley Blackwell

63 POSTER PRESENTATIONS High Poly(adp-ribose) Polymerases1 (parp1) Activity Is Detected In Bubalus Bubalis Milk Somatic Cells From Infected Udder Quarters

Giovanna DE MATTEIS1, Anna REALE2, Maria Carmela SCATÀ1, Francesco GRANDONI1, Vittoria Lucia BARILE1

1Consiglio per la ricerca in agricoltura e l’analisi dell’economia agraria (CREA), Italy 2Sapienza Università di Roma, Italy [email protected], [email protected], [email protected], [email protected], [email protected]

During mastitis the innate immune system responds by the production of ROS that cause oxidative DNA damage and trigger the formation of poly (ADP-ribose) (PAR) by poly(ADP-ribose) polymerases1 (PARP1). Besides DNA repair, PARP1 activity has also been implicated in several cellular processes (apoptosis and necrosis) and in inflammation. Recently, we evaluated PARP1 activation and PAR formation in bovine immune-competent cells suggesting that PARP1 is activated during bovine mastitis. In the present study, we quantified PAR polymers production in quarter foremilk samples from a buffalo cow with absence of clinical sign of mastitis. To monitor PARP activity in milk somatic cells, we used the HT PARP in vivo Pharmacodynamic assay II (Trevigen, USA). In addition, apoptosis of milk somatic cells was monitored using the FITC Annexin V/Dead Cell Apoptosis Kit (ThermoFisher Scientific). Although at time of milk sampling the buffalo did not show clinical sign of mastitis, results of the bacteriological analysis indicated that the right-rear, the left-rear and the left front quarters were infected with S. agalactiae and showed an increased SCC (2,400,000÷7,500,000 cell/mL) respect to quarter that remained bacteriologically negative and with low somatic cell count (SCC; 154,000÷218,000 cells/mL) throughout the monitoring period.

Results showed that PAR amounts in milk leukocytes were significantly higher (P≤0.05) if coming from infected quarters respect to the right-front non-infected quarter (average value of PAR: 580±45.00 pg/mL vs 372±19.6 pg/mL respectively).

Furthermore, the infected quarters had a significant lower level of live and apoptotic cells (23 vs 40%, p≤0.05), and a significant increase of necrotic cells (39 vs 17%, p≤0.05) respect to the healthy quarter.

Overall these results showed that PAR levels were higher in milk somatic cells from infected quarters where an inflammatory response is on course as shown by the increased number of necrotic cells. This suggest PARP1 involvement in udder inflammation and underpins to exploit PAR as potential markers of mastitis.

[1] Assessment of Poly(ADP-ribose) Polymerase1 (PARP1) expression and activity in cells purified from blood and milk of dairy cattle [2] Giovanna De Matteis, Anna Reale, Francesco Grandoni, Mirella L. Meyer-Ficca, Maria Carmela Scatà, Ralph G. Meyer, Luca Buttazzoni, Bianca Moioli [3] Veterinary Immunology and Immunopathology (2018) 202:102-108

64 POSTER PRESENTATIONS Diagnosis of Tubercolosis In Water Buffalo with The Gamma-Interferon Assay: Preliminary Results

Alessandra MARTUCCIELLO1, Nicoletta VITALE2, Piera MAZZONE3, Alessandro DONDO2, Ivonne ARCHETTI4, Fabrizio GAMBERALE5, Maria Lodovica PACCIARINI4, Maria Beatrice BONIOTTI4, Esterina DE CARLO6, Antonio LIMONE7

1National Reference Centre on Water Buffalo Farming and Productions Hygiene and Technologies Istituto Zooprofilattico Sperimenta, Italy 2Istituto Zooprofilattico Sperimentale Piemonte, Liguria e Valle D’Aosta , Italy 3Istituto Zooprofilattico Sperimentale dell’Umbria e delle Marche , Italy 4Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna, National Reference Centre for Tuberculosis , Italy 5Istituto Zooprofilattico Sperimentale di Lazio e Toscana, Italy 6Istituto Zooprofilattico Sperimentale del Mezzogiorno, National Reference Centre for Hygiene and Technologies of Water Buffalo F, Italy 7Istituto Zooprofilattico Sperimentale del Mezzogiorno, Italy [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], esterina. [email protected], [email protected]

The interferon-gamma (IFN-γ) assay is frequently used for diagnosis of Mycobacterium bovis infections in buffaloes [1]. In this study blood was stimulated with two couple of purified protein derivatives (PPDs), avian (PPDA) and bovine (PPDB), from Lelystad (Lel) and from Italy (IT); phosphate buffer saline (PBS = N) was used as a negative control. Three interpretative criteria were used: 1) if Lel and IT PPDB≥2N and PPDB/PPDA≥1.1 then POSITIVE else NEGATIVE if PPDB/ PPDA≤0,9 and inconclusive (IC) for intermediate results; 2) if Lel or IT PPDB≥2N and PPDB-PPDA≥0.050 then POSITIVE else NEGATIVE; 3) if Lel or IT PPDB-N≥0.1 and PPDB-PPDA≥0.1 then POSITIVE else NEGATIVE. Specificity (Sp) was estimated on 466 buffalos from Tuberculosis (TB) free herd while Sensitivity (Se) was estimated on 71 animals from a confirmed TB outbreak as well as latent class model using Bayesian techniques.

Criteria 1 showed 99.1% Sp (CI95% 97.38-99.8%) and 91.5% Se (CI95% 82.5-96.8%), while without IC results the Se was 81.25%. Criteria 2 resulted 96.8% Sp (CI95% 94.5-98.3%) and 86.3% Se (CI95% 76.3-96.7%), while criteria 3 showed 97.3% Sp (CI95% 95.1-98.7%) and 98.1% Se (CI95% 89.7-99.9%). Criteria 1 is the most conservative and for accuracy obtains better performance, but leaves more IC results. Latent class model showed 75% Se for IFN-γ assay higher than SIT (Se 45%). Also the evaluations carried out on the relative Se versus the post-mortem inspection indicate a greater Se of the IFN-γ compared to the SIT. All the animals with lesions (n=11) were positive to IFN-γ, while only one was positive at SIT test. However, these data need confirmation due to the small sample of this study.

[1] E.M.D.L. van der Heijden, Field application of immunoassays for detection of Mycobaterium bovis infection in the African buffaloes (Syncerus caffer), 169:68-73, 2016.

65 POSTER PRESENTATIONS The Prediction of Lactation Milk Yield from Partial Milk Record in Anatolian Water Buffalo

Mehmet Ihsan SOYSAL1, Mehmet AKSEL2, Eser Kemal GÜRCAN1, Emel ÖZKAN ÜNAL1, Serdar GENÇ3

This study was researched relationship between total milk yields and partial milk yields of water buffalo reared Istanbul. The whole data set were obtained as daily milk yield records of water buffalo and fourth lactation number was used in this study. The average lactation period were calculated 216.6 day and the average lactation milk yield were calculated 1360.1 kg. Similarly, the average daily milk yield was calculated 6.26 kg. The relationship between lactation milk yield and additive partial milk yield (30-60-90-120-150-180 day) was analyzed with regression equation. Partial milk yields as independent variables and total lactation milk yield as dependent variable were named and simple and multiple regression analysis methods were used (Y=a+bX and Y=a+b1X1+b2X2). The highest coefficient of determination was identified as the most appropriate model in all models. The correlation coefficient has been found as (r=0.74**) between 30-day partial milk yield and total milk yield and (r=0.88**) between 90-day partial milk yield and total milk yield. Also, the correlation coefficient between total milk yield and 180-day incremental milk yield, has been found as (r=0.94**). The relation between total milk yield and additive partial milk yield was analyzed with regression equation. The most suitable regression equation was chosen according to determination coefficient. The determination coefficients (R2) were calculated for 30 and 60 days, using of two independent variables together was 0.77 and using one independent variable (30, 60, 90, 120 and 150 day) partial yields were 0.54, 0.69, 0.77, 0.82, 0.84 and 0.88 respectively. As a result, the partial milk yields could be used to estimate lactation milk yield previously as effectively.

Istanbul Water Buffalo Breeders Association

[1] Soysal Mİ, Küçük R (1996). Siyah Alaca Süt Sığırlarında Eklemeli Kısmi Verimler İle Birinci Laktasyon Süt Veriminin Tahmin Olanakları Üzerine Bir Araştırma. Tekirdağ Ziraat Fakültesi Dergisi S.284-293

66 POSTER PRESENTATIONS Using Animal Need Index to Investigate Welfare Level of The Water Buffalo Farms in Marmara Region

Pembe Dilara AKIN1, Hülya YALÇINTAN1, Nurşen ÖZTÜRK1, Mehmet Ihsan SOYSAL2, Mustafa KÜÇÜKKEBABÇI3, Ömür KOÇAK1

1İSTANBUL ÜNİVERSİTESİ-CERRAHPAŞA, Turkey 2Tekirdağ Namık Kemal Üniversitesi, Turkey 3Sheep Breeding Research Institute, Turkey [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

Water buffalo breeding is a growing field in Turkey because of the increase in consumer demands for special buffalo milk products such as milk, milk cream, mozzarella and ricotta cheese. In order to increase milk yield, some farmers apply intensive rearing system instead of traditional method which is based on an extensive production with poor feeding and management conditions. On the other hand, the vast majority of Buffalo population in Turkey consist of Anatolian Water Buffalo which is an indigenous breed with low milk yield, while is well adapted to the harsh environment, poor breeding and management conditions and also resistant to diseases. Animal Needs Index (ANI) method was developed for evaluation of on-farm welfare levels of dairy cattle. The aim of the study was investigating the possibility of using this method for evaluating the welfare levels of buffaloes. In the current study, breeding conditions of forty-two buffalo farms in Marmara Region were evaluated in terms of animal welfare with ANI method. The enterprises to be included in the study were divided into 3 sub-groups according to herd sizes in three cities of Marmara Region (Istanbul, Kocaeli and Balıkesir), which have the highest numbers of buffaloes in the Region. The sub-groups were formed as; a) Small-scale farms (n: 14): Number of female buffaloes between 10 to 40 heads, b) Medium-sized farms (n: 14): Number of female buffaloes between 41 and 60 heads, c) Large-scale farms (n: 14): Number of female buffaloes over 61. The farms in the study were visited in both summer and winter by four people experienced in ANI assessment. ANI welfare assessment on designated farms were done in five different categories: i) locomotion, ii) social interaction, iii) flooring, iv) light and air and v) stockmanship. Scoring were done for a total of 30 criteria in these five different categories.

The effects of both season and city on total ANI scores of farms were investigated and found significant as a result of the study. The farms in both cities had higher scores in the summer than winter, because animals had access to pastures and yards. In the meanwhile, the farms in Kocaeli had higher scores than the ones in Istanbul for both seasons, because they reared the buffaloes in pasture and yards for longer periods (both hours/day and day/year). In concluaion, ANI evaluation system can be adapted for water buffaloes, but it will be an inadequate method for behaviour and health evaluation at farm level, which are really important for welfare of an animal. However, it can be used as a starting point for developing more suitable welfare evaluation systems for water buffaloes.

The results of the study might be used for developing a new method for evaluating buffaloes’ welfare levels in the current breeding conditions and at farm level.

We would like to thank to Republic of Turkey Ministry of Agriculture and Forestry General Directorate of Agricultural Research and Policies, Şenol Çakır, Talat Tan, Mehmet Aksel and Rabiya Birkiye.

67 POSTER PRESENTATIONS Magnetic Resonance Imaging Features of Brain Lesions From Penetrative Stunning of River Buffaloes Slaughtered In Brazil

Ariane DANTAS1, Jéssica Leite FOGAÇA1, Michel De Campos VETTORATO2, Viviane Maria CODOGNOTO1, Stella Maris Teobaldo TIRONI1, Andressa Filaz VIEIRA1, Suzane BROCHINE1, Vânia Maria De Vasconcelos MACHADO1, Eunice OBA1

1São Paulo State University (Unesp), School of Veterinary Medicine and Animal Science, Botucatu, Brazil2 Unesp - FMVZ, Brazil [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

Stunning is performed to make animals unconscious and to render them insensitive to pain prior to slaughter. However, in buffaloes, certain anatomical features of the head make effective stunning a more complex task, when compared to bovine. The objective of this study was to describe magnetic resonance imaging features of brain damage caused by local trauma due to stunning. Eleven heads of Bubalus bubalis females from 15 to 30 months of age slaughtered in the same slaughterhouse and previously stunned by a single professional using a 7kg 304 stainless steel penetrative pistol, with working pressure of 182psi, production capacity of 250 animals/ hour. Heads were kept frozen until 48 hours before the examination. After complete thawing, magnetic resonance imaging was performed using 0.25 Tesla magnetic field equipment attached to a radio frequency coil (1 frequency channel). The sections were: Spin Eco T1 (SE T1) and Fast Spin Eco T2 (FSE-T2), both in the sagittal (FOV = 26cm) and transverse (FOV= 24cm) and FAST FLAIR in the transverse plane (FOV= 24cm). The images obtained were analyzed using Synapse Pacs® software. A degree of brain laceration, which corresponded to 0 (without detectable lesion), 1 (moderate lesion, anatomical structures still identifiable) and 2 (severe lesion, very compromised anatomical structures) were evaluated. A bleeding hyposignal score was attributed, being 0 (absent) and 1 (present). Volume of intracranial air (cm3) and number of shots of pistol and bones struck in each head were assessed. Laceration grade 2 (45.45%) and 3 (36.36%) and hemorrhage were detected in most of the pieces (81.81%). Average intracranial pressure was 82.76cm3. Occipital perforation occurred in 100% of the heads. In those which received 2 or 3 shots, frontal bone was also perforated . Severity of brain lesions, loss of vascular integrity and variation of intracranial pressure was proportional to the intensity of external trauma. Obtaining images and data of this kind of neurological traumatism is key for the establishment of efficient stunning methods for buffaloes in Brazil.

68 POSTER PRESENTATIONS Consumers’ Perception and Awareness of Buffalo Mozzarella Authenticity

Andressa DA COSTA LIRA THOMAZ DE AQUINO, Evelise OLIVEIRA TELLES

University of São Paulo, Brazil [email protected], [email protected]

This study aimed to investigate consumers’ perception and awareness of buffalo mozzarella authenticity. Questionnaire was used to address motivation for buying it, consumers’ perception of its authenticity, label reading, cheese fraud, willingness to pay for authentic product, and trust in control agencies. Interviews were carried out in supermarkets, Mooca, Municipality of São Paulo city, Brazil. Interviewees must be buffalo mozzarella consumer and to be of age. Sample was stratified at “high level of knowledge” when they revealed knowledge about cheese fraud and “low level” when they did not or presented misconceptions. The methodology for discourse analysis was the Discourse of the Collective Subject, (LEFEVRE; LEFEVRE, 2005). 5-point Likert Scale was used for analyzing trust in the control agencies; Mann-Whitney and Fischer tests for identifying associations of the parameters with “knowledge about cheese fraud”, Kruskal-Wallis test for assessing associations of “confidence level” and “educational level”. 120 interviewees’ answers (80% female, 75% higher education, 49 years old mean age) showed that the main motivation for buying buffalo mozzarella was organoleptic characteristics and, although 41% reported nutritional motivation, knowledge misconceptions were observed; 82% recognize it by the presentation (shape and color) and confidence in brand and commercial establishment. They trust in institutions for fraud prevention, but they do not seek their seals. 83% read label, but 8% read “Ingredient List” (4% for avoiding the purchase of a wrong product by mistake). There is little knowledge about buffalo mozzarella fraud, although the mixture of different milks was considered fraud. There is willingness to pay for the authentic product, despite the misunderstanding about its nutritional characteristics. Stratification of sample did not influence discourse (p> 0.05). Therefore, consumer do not have knowledge about this fraud to take measures to mitigate the risk of being cheated, nor what measures are within their reach.

This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001

[1] LEFEVRE; F.; LEFEVRE, A. M. C. Discurso do sujeito coletivo: um novo enfoque em pesquisa qualitativa (desdobramentos). 2. ed. Caxias do Sul, RS: EDUCS, 2005.

69 POSTER PRESENTATIONS Determination of The Cryoscopic Index in Mediterranean Buffalo Milk: Identification of The Reference Parameters, Confirmation of The Adulteration and Calculation of The Amount of Water Added.

Pasquale FRAULO, Domenico VECCHIO, Esterina DE CARLO

National Reference Centre on Water Buffalo Farming and Productions Hygiene and Technologies Istituto Zooprofilattico Sperimenta, Italy [email protected], [email protected], [email protected]

Although the adulteration by adding water in milk is considered a “outdated fraud”, in reality, in the production area of the “DOP Campania buffalo mozzarella” in Italy, it is still a problem of considerable importance, causing not a few controversies between the farmers and dairies . The problem lies in the fact that, to detect adulteration in buffalo milk, the cryoscopic index is used, which has no threshold value and is extremely variable between farms and, on the same farm, varies according to the lactation period. The authors conducted a study on about 20 buffalo breeding farms, controlled with mass fortnightly milk sampling and analysis of the main milk quality parameters for about a year, and 24 buffalo breeding farms controlled with monthly individual milk sampling, to identify a threshold value of a cryoscopic index to be used for detecting the suspicion of adulteration of Mediterranean buffalo milk. The authors also identified a diagnostic algorithm for the confirmation of adulteration of buffalo milk and a calculation system for the quantification of extraneous water added fraudulently in milk, by tests carried out on milk with a known title artificially adulterated. The experimentation highlighted a significant correlation between the fat concentration and the cryoscopic point of buffalo milk, suggesting the use of a system for calculating the amount of water added, which would consider the variation of fat concentration in milk (% H2O ext. = (B -T) x (100-S) / B). The proposed added water calculation formula has shown greater accuracy than the commonly used calculation systems, with differences from the expected value of up to 2%. The authors also elaborated the percentile values of the freezing point, identifying the ninetieth percentile as -0.520 ° C for mass milk as the threshold value for suspected adulteration of milk. As a confirmation of the adulteration of buffalo milk, the authors verified the validity of the “stall test”, which consists in performing a significant sampling of individual milk and in calculating the geometric mean of the freezing point.

[1] Alais C., 2010 Scienza del latte: principi di tecnologia del latte e dei derivati III edizione ed. Tecniche Nuove [2] Pesce A., Salzano C., de Felice A., garofalo F., Liguori S., De Santo A., palermo P., Guarino A., 2016. Monitoring the freezing point of buffalo milk. Italian Journal of Food Safety 2016; 5:5691. 95-97

70 POSTER PRESENTATIONS Buffalo Milk Produced in Lazio Region (Italy): Hygienic, Milk Quality and Milk Clotting Properties During One Year of Study

Carlo BOSELLI1, Antonio BORGHESE2, Laura GEMMA1, Giuseppe BITONTI1, Riccardo BICOCCHI1, Alessandro PROIETTI1, Gilberto GIANGOLINI1, Marcella GUARDUCCI1, Simonetta AMATISTE1

1Istituto Zooprofilattico Sperimentale del Lazio e della Toscana M. Aleandri, Italy 2International Buffalo Federation, Italy [email protected], [email protected], [email protected], [email protected], riccardo.bicocchi@ izslt.it, [email protected], [email protected], [email protected], [email protected]

In Italy the buffalo population is around 400,792 heads (2017), the milk production is important for economy because it is used to produce different cheeses mainly represented by Mozzarella di Bufala Campana PDO (EC Regulation n 1107/96). In European Union, EC Regulation n 853/04 establishes the limit of Total Bacterial Count (TBC) for bulk tank milk (1,500,000 cfu/ml).

The aim of this study was to analyze the TBC, milk quality parameters, somatic cell count and milk clotting properties in 2,027 milk bulk samples collected during entire 2018.

The geometric mean of TBC (Bactoscan) was 313,042 cfu/ml.

The average value of chemical parameters (CombiFoss) were: fat 7.87%, protein 4.50%, casein 3.69%, lactose 4.77%, solid non fat 10.48% and somatic cell 181,000 cell/ml. The main technological traits (Formagraph) were: rennet coagulation time (13.61 min.), curd firmness at 30 minutes (47.59 mm) and at 60 minutes (50.61 mm). TBC was significantly correlated with rennet coagulation time (r = -0.24), and curd firmness at 60 minutes (r = -0.19). Considering the legal limit for total bacterial counts, 331 (16.33%) samples had counts exceeding 1,500,000 cfu/ml. The milk samples were divided in two groups, above and below the regulatory limit of TBC and put in relation with the main technological traits. Results of statistical analysis (ANOVA) showed that the two TBC level significantly influenced only rennet coagulation time and curd firmness at 60 minutes. The TBC group below the legal limit, showed a longer rennet coagulation time (14.12 vs 11.20 min) and curd firmness at 60 minutes (51.79 vs 46.22 mm).

In Italy, the dairy industries apply the milk payment system on the basis of the chemical composition and TBC. TBC is the main responsible of the better price and indicator of hygienic and cleaning conditions in milk production and handling in the farm, as well as of its adequate storage and refrigeration.

[1] Borghese A. 2013 . Buffalo livestok and products in Europe. Buffalo Bull., 32 (Special Issue 1): 50-74. [2] Giangolini G., Amatiste S., Proietti A., Boselli C., Pietrini P., Rosati R., (2003). Evaluation of Bactoscan FC for determination of total bacterial count in buffalo raw milk; comparison with Bactoscan 8000. Atti II Congresso Nazionale sull’Allevamento del Bufalo, 137-146. [3] Pasquini M., Trombetta M. F., (2014). Microbiological quality and safety of Mediterranean buffalo bulk milk. 7° Convegno Nazionale ARNA, 2-4 October, Cagliari, 45-47. [4] Regulation (EC) No. 853/2004 of the European Parliament and of the Council of 29 April 2004 laying down specific hygiene rules for food of animal origin. Off. J. Eur. Commun. L, 139 (2004), pp. 55–205 [5] Tripaldi C., Amatiste S., Giangolini G., Palocci G., Borghese A. Milk Quality and Product. In Borghese and C.R.A. Editor. Buffalo Livestock and Products, Roma (2013), 8:232-259. [6] Carlo BOSELLI, Gilberto GIANGOLINI, Francesco FILIPPETTI, Tarcisio TULLIO, Antonio BORGHESE, Simonetta AMATISTE, Remo ROSATI. (2015). Quality, safety and seasonal aspects of Buffalo milk produced in Lazio Region (Central Italy). Proceedings of 8th Asian Buffalo Congress, Istanbul - Turkey.

71 POSTER PRESENTATIONS Determination of The Best Lactation Curve Model and Lactation Curve Parameters Using Different Nonlinear Models for Anatolian Buffaloes

Ertuğrul KUL

Department of Animal Science, Faculty of Agriculture, University of Kırşehir Ahi Evran, Kırşehir, Turkey [email protected]

Abstract The present investigation was conducted to determine lactation curve parameters and to choose the best model that defining the lactation curve. Data were composed of 4897 test day milk yield (TDMY) records from Anatolian buffaloes calved during 2014-2017 raised under different farm conditions in Amasya Province, Turkey. Five different lactation curve models as Wood (WOD), Cobby and Le Du (CLD), Exponential (EXP), Parabolic Exponential (PEXP) and Quadratic (QUA) models were used. The Adjusted multiple coefficient of determination (R2adj) and the residual standard deviation (RSD) were evaulated as a parameter to detect the best fitted lactation curve model. The results revealed that WOD model described the highest R2adj (0.98) and with the lowest RSD (0.065), besides the highest R2adj (0.97) and the lowest RSD (0.087) were also observed in CLD model.

To conclude, WOD and CLD models were detected to be the most suitable models defining the lactation curve of Anatolian buffaloes. As a result, using the parameters detected via Wood and Cobby and Le Du models in breeding studies will contribute significantly to the researches in this direction in these herds.

This research was supported financially by Republic of Turkey, Ministry of Agriculture and Foresty, General Directorate of Agricultural Research and Policies. We would like to thank Aziz Şahin and Amasya Water Buffalo Breeders’ Association for valuable technical assistance.

72 POSTER PRESENTATIONS Market Structure and Marketing Problems Of Buffalo Milk And Milk Products In Kayseri Province, Turkey

Arş. Gör. Oğuz ALTIN, Prof. Dr. Yılmaz ARAL

Ankara Üniversitesi Veteriner Fakültesi, Turkey [email protected], [email protected]

It was aimed to determine the market structure and marketing problems of buffalo milk and its products by conducting a survey with 29 buffalo breeders in the province of Kayseri. The mean age of breeders 47 and the average number of buffaloes per enterprise was reported as 33. The breeders stated that 72.4% of them had primary and lower education. The breeders also reported that they frequently market their products as yoghurt (cow-buffalo mix) and then cream, milk, butter, pure buffalo yoghurt and cheese, respectively. Some factors such as consumer demands, regional population, age of producers, scales of buffalo enterprises have been effective in the variety of marketed milk-milk products.

When breeders market their products, they often prefer to reach consumers without any intermediary, which is a direct marketing channel. Among indirect channels, sale through the supermarket/deli is the most preferred channel. Afterwards, it is preferred to market buffalo milk and products to factories, breakfast restaurants, restaurants, dairies and wholesalers respectively. It is important to ensure the quality price relationship. Due to the lack of a standard value of cow-buffalo milk mixture ratios, unfair competition occurs between producers, this situation also has negative consequences for consumers. Collection and processing of buffalo milk in certain areas will contribute the producer’s income levels and to ensure sustainability in production. In order to protect the rights of producers and consumers, labels such as brands and geographical indications will be effective. In this context tag which contain the contents of the products, production and expiry dates, production sites etc. information will be useful. In a production area, where most of the enterprises are disorganized and small-scaled, its difficult for each enterprise to adopt its own brand both financially and in compliance with the legislation. These and similar problems can be solved with an effective cooperative model.

73 POSTER PRESENTATIONS Effect of Supplementing Acacia Arabica Bark Dry Extract to Total Mixed Ration on The Performance of Lactating Buffaloes

Haneet Kaur HANJRAH1, Manju WADHWA1, Jaspal Singh HUNDAL1, Mohinder Pal Singh BAKSHI1, Amit SHARMA1, Puneet MALHOTRA2, Baljinder Kumar BANSAL2

1Department of Animal Nutrition, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana- 141 004, India 2Directorate of Livestock Farms, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana- 141 004, India [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

This study was taken up to assess the effect of supplementing Acacia arabica (Babul) bark to the total mixed ration (TMR) on the performance of lactating buffaloes. A TMR containing concentrate mixture, Trifolium alexandrium (Berseem) green fodder and wheat straw in 40:18:42 ratio on DM basis was formulated. TMR supplemented with or without A. Arabica @ 1% was evaluated by in-vitro gas production technique and on 12 lactating buffaloes, divided into 2 equal groups fed for 180 days. Before the termination of feeding trial, a 14-day digestion trial was conducted by indicator method using chromic oxide green as an external marker. On the last day of trial, blood samples were taken from each animal by puncturing jugular vein. The enteric methane production was assessed by SF6 tracer technique. The data were analyzed by one way ANOVA. Supplementation of TMR with A. arabica bark did not affect the net gas production, ME content and digestibility of nutrients; improved the VFAs production and fermentation efficiency; decreased (p<0.05) methane production resulting in higher microbial biomass synthesis. Supplementation of diet with Acacia arabica bark did not affect DM intake, but digestibility of CP, EE and cellulose in buffaloes was improved (p<0.05) on supplementation of TMR with A. arabica bark. The methane emission was decreased (p<0.05) by 37% in the TMR supplemented with A. arabica bark. Supplementation of diet with Acacia arabica bark did not show any adverse effect on milk yield; improved (p<0.05) milk fat percent, however, protein and SNF percent decreased. The feed conversion efficiency expressed as FPCM/DM intake was improved in buffaloes fed diet supplemented with A. arabica bark. TMR supplemented with A. arabica bark decreased (p<0.05) saturated fatty acids, increased mono unsaturated fatty acids and poly unsaturated fatty acids in milk fat. It was concluded that supplementation of TMR with Acacia arabica bark mitigated enteric methane emission, without affecting milk yield, but milk composition was altered. ICAR (Indian Council of Agricultural Research)

74 POSTER PRESENTATIONS Increase in Nitrogen Retention in Buffaloes with Diets Based on Whole Soybeans And / Or Antioxidants

Lucia Maria ZEOULA1, Erica MACHADO1, Bruna Calvo AGUSTINHO1, Jocasta CARRARO1, Silvia Cristina AGUIAR2, Nadine Woruby SANTOS1, Emerson Henri YOSHIMURA3

1State University of Maringá, Brazil 2University of the State of Mato Grosso, Brazil 3Universidade Estadual de Maringá, Brazil [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

There are several studies that evaluate the role of secondary plant metabolites, such as polyphenols, essential oils and saponins, in microbial fermentation in the rumen. The aimed of this study was to evaluate microbial protein synthesis of buffalo whose the diets were supplemented with ground whole grain soybeans and antioxidants. Four rumen cannulated female crossbread buffaloes (506 kg ± 29 kg of body weight), were distributed in a Latin square design (4x4) and orthogonal contrasts were used to compare the effects of treatments. Four diets were tested: control (with soybean meal); GS (diet with ground whole soybeans); GSYM (GS diet + yerba mate (YM), 30 g/kg DM), and GSYME (GSYM diet + vitamin E, 375 UI/ kg DM). The control diet had 23 g/kg DM of ether extract, and diets supplemented with ground whole grain soybeans had 53.3 g/kg DM of ether extract. The microbial nitrogen synthesis compounds in the rumen (Y, g N/day) was estimated based on absorbed purines (X, mmol/day), according Chen and Gomes (1992). The addition of ground whole grain soybeans to buffalo diets increased microbial production, and efficiency of microbial protein synthesis when compared to control (111.32 vs 127.95 g microbial CP/kg ruminal digestibility of organic matter (DOMR)). The yerba mate also improved the efficiency of microbial protein synthesis (127.95 vs 140.49 g microbial CP/kg DOMR). The effect was potentially increased by the addition of YM and by combined antioxidants, YM and vitamin E (126.95 vs 154.03 g of microbial CP/kg of DOMR). The results suggest that the microbial protein synthesis can be indirectly influenced by lipids, reducing the protozoa population and increasing the production of microbial nitrogen. The effects of YM on microbial protein synthesis may have occurred due to the saponins of YM, which also reduces the rumen ciliate protozoa population. The use of YM and vitamin E combined the protective action of vitamin E on cell membranes with action of YM antioxidants. Thus, it is concluded that the increase of retention of nitrogen in the rumen can be due to the use of dietary lipids and antioxidants.

[1] Chen, X.B., Gomes, M.J. 1992. Estimation of microbial protein supply to sheep and cattle based on urinary excretion of purine derivatives- an overview of technical details. International Feed Research

75 POSTER PRESENTATIONS Superovulatory Response and Embryo Recovery Following Ablation-induced Follicular Wave Synchronization in Buffalo (bubalus Bubalis)

Mrigank HONPARKHE1, Vinod GANDOTRA1, Joga Singh MATHAROO1, Dinesh DADARWAL2, Sarvpreet Singh GHUMAN1, Parkash Singh BRAR1, Jaswant SINGH2

1Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, India 2University of Saskatchewan, Saskatoon, Canada [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

Our objective was to evaluate the superstimulatory response, ovulation rate, luteal function, and recovery rate of transferable embryos in water buffaloes when follicular stimulating hormone (FSH) treatment was initiated at the start of a follicular wave induced by ablation of the extant dominant follicle. We compared the luteal function after single ovulation in unstimulated wave versus multiple ovulations after FSH-stimulation. Murrah buffaloes (n=6) were given prostaglandin F (PGF)-2α analog (Cloprostenol 500μg, i.m.) to induce luteolysis and timing of the spontaneous ovulation was detected by daily ultrasonography.Transvaginal ultrasound-guided follicle ablation procedure was performed on Days 7 or 8 post- ovulation and the FSH treatment was initiated 30 h later (50 mg i.m. x 8 doses at 12 h intervals). Buffaloes were administered two PGF injections at 72 and 84 h, luteinizing hormone (LH, 25 mg, i.m.) at 108 h and were inseminated at 120 and 132 h. Following LH injection, ovarian ultrasound was performed at 12 h intervals for 4 days to detect ovulations.Embryos/ ova were recovered on Day 5 after the first insemination. Data were analyzed by SAS using Proc Mixed analyses.The higher (P<0.05) number of 6-8 mm (on Day1-3 post wave emergence) and >8 mm (on Day 4-6 post wave emergence) size follicles were observed in superstimulated wave compared to the unstimulated wave. Based on the disappearance of ≥ 8mm follicles and presence of CL on Day 4 after LH the ovulation rate was 61.5 % and 74.3 %, respectively. Mean (±SEM) progesterone concentration and luteal area on Day 4 post-ovulation were greater (P<0.05) after superovulation as compared to single ovulations (1.1-5.5 ng/ml vs 0.5-0.7 ng/ml and 1076.1±180 mm2 vs 135.1±14.7 mm2). Out of total 3.0±0.8 embryo, 73% (2.2±0.2) were of transferable grade. In conclusion, initiation of superstimulatory treatment subsequent to the synchronization of follicular wave emergence using transvaginal ultrasound-guided follicular ablation enabled to improve superovulatory response and transferable embryo recovery in Murrah buffaloes.

76 POSTER PRESENTATIONS Artificial Insemination Out of Breeding Season in Water Buffaloes in Argentina

José KONRAD1, Ramon CAMELINO2, Roberto YUPONI2, Natalia VALLEJOS2, Alexis BANDEO2, Jesus BERDUGO3, Gustavo CRUDELI2, Pablo MALDONADO VARGAS2

1Facultad de Ciencias Veterinarias, UNNE. CONICET, Argentina 2Facultad de Ciencias Veterinarias, UNNE, Argentina 3Grupo BIOGEM, Universidad Nacional de Colombia, Colombia [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

Seasonality in the reproduction of water buffalo is one of the main problems of breeders to have a constant production. Some authors have been suggested that the use of out of breeding season ovulation induction programs, might help to overcome this problem. The objective of this work was to evaluate the effect over pregnancy rates the application of an ovulation synchronization protocol designed for out of breeding seasons. This work was performed in September of 2018, during out of breeding season in a farm located in Corrientes, Argentina. 26 adult normal females of similar weight and parity were synchronized, using progesterone, eCG, prostaglandin and estradiol cipionate protocol. Ten days before the starting the synchronization protocol half of the animals were treated with melatonin implants at a dose of 18mg/50 kg of live weigh. At the beginning of the protocol blood sample to determine antimüllerian hormone (AMH) was taken. Inseminated by the same technician with frozen semen of Murrah bulls of proved fertility. Pregnancy rates were compared with those obtained during breeding season artificial insemination protocols in the same farm. Data were analyzed using comparison of proportions, using Tukey test and statistical difference were considered p>0.05. 9 of the 26 females became pregnant (34.61%) compared to (52.5%) during breeding season, with no statistical difference (p=0.2076). No difference in AMH levels were found between pregnant and non-pregnant females, 245.5 vs 238.7 pg/ml (p=0.1496). No effect of male in pregnancy rates were observed. This result shows comparable results in terms of pregnancy rates in out of breeding’s season allowing farmers to implant this strategy as part to satisfy market demand´s. As reported by others they are no effect of ovarian reserve analyzed through AMH levels over pregnancy rates.

[1] Ghuman SPS. Singh J. Honparkhe M. Dadarwal D. Dhaliwal GS. Jain AK. 2010. Induction of ovulation of ovulatory size non-ovulatory follicles and initiation of ovarian cyclicity in summer anoestrous buffalo heifers (Bubalus bubalis) using melatonin implants. Reprod Dom Anim 45: 600–607. [2] Kavita. Phogat JB. Pandey AK. Balhara AK. Ghuman SS. Gunwant P. 2018. Effects of melatonin supplementation prior to Ovsynch protocol on ovarian activity and conception rates in anestrous Murrah buffalo heifers during out of breeding season. Reprod Biol. 18: 161-168. [3] Konrad JL. Garrido MJ. Maldonado Vargas P. Crudeli GA. 2016. Effect of administration of equine chorionic gonadotrophin after artificial insemination on pregnancy rates in buffalos in Argentina. 11th World Buffalo Congress, Cartagena, Colombia. P: 19. [4] Neglia G. Capuano M. Balestrieri A. Cimmino R. Iannaccone F. Palumbo F. Presicce GA. Campanile G. 2018. Effect of consecutive re-synchronization protocols on pregnancy rate in buffalo (Bubalus bubalis) heifers out of the breeding season. Theriogenology. 113: 120-126. [5] Redhead AK. Siew N. Lambie N. Carnarvon D. Ramgattie R. Knights M. 2018. The relationship between circulating concentration of AMH and LH content in the follicle stimulating hormone (FSH) preparations on follicular growth and ovulatory response to superovulation in water buffaloes. Anim Reprod Sci. 188: 66-73.

77 POSTER PRESENTATIONS Age of First Calving Season and Calves Growth Data in Anatolian Water Buffaloes In Çorum Province

Mehmet Ali YILMAZ1, Engin ÜNAY1, Yusuf KAPLAN2

1International Center for Livestock Research and Training, Lalahan, Ankara, Turkey 2General Directorate of Agricultural Research and Policies, Ankara, Turkey [email protected], [email protected], [email protected]

In this study, it was aimed to determine the relationship between age of first calving and season of Anatolian Water Buffaloes (AWB) and the birth weight (BW), 6 month weight (SMW) and yearling weight (YW) of their calves. For this purpose, the data belonging 137 primiparous AWB and their calves in the scope of National Animal Breeding Project in Public Conditions carried out by General Directorate of Agricultural Research and Policies were utilized in the study. Çorum province, which is located in Middle Anatolia, has 800 m altitude and large geography and different ecological areas with an area of 12.820 km2. It is understood from the findings of the historical excavations that the AWB population was domesticated centuries ago. Traditionally, cultivation of AWB in the area is made semi intensively depending on pasture. Breeding Project İn Farm has been carried out in Çorum province since 2012. Mating usually occurs in springs and summers. In 2019, the number of cows and bulls in the project was 826 and 60, respectively. In this study, the BW, SMW and YW of the calves of 137 primiparous AWB in the project were evaluated. 78 (%56.93) numbers of 137 primiparous AWB gave birth in spring, while 25 (18.24%) gave birth in the summer months, 12 (8.75%) gave birth in autumn and 23 (16.78%) gave birth in winter. In this region, it is observed that more than half of the first calves are serviced in spring. In the scope of the study, the birth weight six months and one age live weights were found for the spring; 29.86 kg, 125.16 kg, 167.88 kg, summer; 30.49 kg,123.53 kg,172.52 kg, autumn; 28.61 kg, 124.25 kg,160.49 kg and winter; 30.96 kg, 126.18 kg, 176.35 g, respectively. The BW of young buffalo calf, seasonal effects of SMW and YW live weight were not significant (P>0.05). As a result of our study, when the data were evaluated, the insignificance of seasonal effects was not consistent with the literature. It is considered that it would be appropriate to evaluate the subject by using data of more animals in order to understand the effect of seasons more clearly.

This project is funded by the Ministry of Agriculture and Forestry, under the coordination of the Directorate of Livestock and Aquaculture Research, General Directorate of Agricultural Research and Policy, in cooperation with Corum Provincial Breeders’ Breeders Association.

78 POSTER PRESENTATIONS Relationships of Body Hygiene Scores and Udder Surface Temperature with Milk Somatic Cell Count and Milk Components in Anatolian Buffaloes

Ayşenur BULU, Hüseyin ERDEM

Ondokuz Mayıs Üniversitesi Ziraat Fakültesi, Turkey [email protected], [email protected]

The objective of this study was to reveal the effects of dirtiness level of udder, foot and rear legs on somatic cell count (SCC) and components of milk in Anatolian buffaloes. A total of 75 buffalo cows reared in five farms located in Bafra district of the Black Sea region and their milk samples were constituted to be the study materials. Udder (UHS), rear legs (LHS) and foot (FHS) hygiene scores and udder surface temperature (UST) values were recorded monthly. For scores, a scale with 1 to 4 points where higher points reflect dirtier was used. To SCC and milk analyses, 50 ml milk samples were collected from each animal in test days. Total dry mater (TDM), fat (F), non-fat dry mater (NFDM), protein (P), lactose (L), mineral (M), density (D) and freezing point (FP) tests were applied for determining raw milk composition. The highest scores for all body parts were recorded in spring and summer seasons. While sampling season affected UHS, FHS and UST (P˂0.001), the dirtiness was observed in spring and summer. In contrast, the highest UST values were recorded in winter and autumn. Increase in UHS caused to drops in DM, F, NFDM, P and D, to increase in FP. Buffalo cows with 4 points had lower levels in milk components (P<0.001). The LHS and FHS had negative effect on DM, NFDM, P, FP and D. Besides, weak correlation coefficients were estimated between SCC and UHS, LHS and FHS (r=0.037, 0.052 and 0.016, respectively). It was concluded that dirtiness levels of three body parts of the cows were negatively affected some milk components, but SCC was not affected by hygiene scores or UST.

79 POSTER PRESENTATIONS The Effects of Water Buffalo Improvement Project Conducted in Public Farms of Samsun Province On Breeding

Nazlı SAYAR YAPAR, Hüseyin ERDEM

Ondokuz Mayıs Üniversitesi Ziraat Fakültesi, Turkey [email protected], [email protected]

“National Improvement Project on Buffaloes Reared at Village Conditions” a breeding project which has been carried out in 18 cities under coordination of General Directorate of Agricultural Researches and Politics of Turkey Ministry of Agriculture and Forestry to elevate yield and revise husbandry conditions. This study was aimed to reveal the effects of the project on buffalo breeding in Samsun province. Data of the study were obtained from questionnaires collected by interviews from a total of 48 breeders chosen from 100 breeders by simple random sampling method. Of the farms, 27.8 % had ≥100 (head) and 25% had < 20 (head) animals. It was noted that 62.5% of the breeders kept the records on milk and buffalo calves, and also, 35.4% of those kept milk yield, calf birth and weight records as soon as introducing to the project. Similarly, it was determined that 85.4% of the breeders have milked animals since the project; however, 10.4% of those have milked their animals after beginning this project. Raw milk was highly (70.2%) sold in the local bazaars, and the percentage of the persons, who thought that the project was effective on milk marketing, was 59.6%. While 72.9% of the breeders changed the feeding regime with project, concentrate using rate with project was recorded to be 32.6%. Of the breeders, 97.9% was satisfied from the supports as part of the project, but 47.9% of those pointed out to inadequacy of the supports. The rates of persons thought the project contributed to buffalo rearing and it was effective on breeder selection were 93.7% and 95.8%, respectively. However, the percentages of the breeders, who emphasized that the project improved milk production, animal number and husbandry conditions, were determined as 34.9%, 34.9% and 60.5%, respectively. While the percentage of the breeders those used the supports of the project was calculated as 95.8, 97.9% of the breeders was willingly for continuing it.

80 POSTER PRESENTATIONS Evaluation of Some Spermatological Characteristic of Different Doses of Frozen Sperm in Anatolian Buffalos

Derya ŞAHİN1, Ilktan BAŞTAN1, Fırat KORKMAZ1, Seher YIRTICI1, Ufuk KAYA2, Hüseyin KİNET1

1Uluslararası Hayvancılık Araştırma ve Eğitim Merkezi Müdürlüğü, Turkey 2Ankara University, Faculty of Veterinary Medicine, United States [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

Anatolian Buffalos is a domestic water buffalos, which is known as Mediterranean Buffalos included in river group in Turkey. In recent years, many scholars concern about spermatological features of Anatolian buffalos. Furthermore, since 2017 in Turkey the first commercial Anatolian Buffalos sperm has been produced in department of artificial insemination laboratory in International Livestock Research and Training Center was sold to various province after checked and qualified by veterinarian. Before the commercial sale stage, we have done producing of frozen semen for trial. For this purpose, two chief Anatolian Buffalos, which have 3-4 years old, was used and a total of 32 ejaculates were taken with artificial vagina. These ejaculates were divided into three groups at different doses: Group-1 (60 x106 sperm/ml), Group-2 (100 x106 sperm/ml), Group-3 (140 x106 sperm/ml). Concentration of fresh semen was examined with spectrophotometric way (Accucell, IMV, France), and motility of fresh semen was evaluated by using a phase-contrast microscope with heating table then it was diluted with AndromedÒ (Minitube, Germany) extender, loaded into 0.25 ml french straws. After the equilibration (+4 0C), it was frozen with a digital freezing device (Digitcool 5300 ZB 250; IMV, France). The frozen semen samples were stored in liquid nitrogen tanks at -196 0C until thawing. Thawing at 36 0C 30 sec in the water bath, spermatozoa motility and kinetic parameter analysis with IVOS I CASA (Hamilton Thorne Inc., Beverly, USA) and the viability spermatozoa (Plasma membrane and acrosomal integrity (PMAI) analysis) were analysed by flow cytometric metod (CytoFLEX, Beckman Coulter, USA). While there was statistically significant differences PMAI between the groups, but no differences was found in terms of kinetic parameters. As a result, in this study, which was carried out belonging to the Anatolian buffalo bull, it was determined that the freezing of semen samples at different doses influenced PMAI.

81 POSTER PRESENTATIONS Effect of Treatment with GNRH At the Moment of Embryos Transfer on The Follicular Dynamics and Conception Rate of Water Buffalo Recipients

Damıana CHELLO1, Julio Cesar BARBOZA DA SILVA2, Guilherme PUGLIESI3, Nelcio Antonio TONIZZA DE CARVALHO4, Julia GLEYCI SOARES3, Pietro SAMPAIO BARUSELLI3

1Department of Animal Reproduction - FMVZ / USP, Italy 2Faculty of Jaguariúna - FAJ , Brazil 3Department of Animal Reproduction - FMVZ / USP, Brazil 4Unit of Research and Development of Registro – APTA, Brazil [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

The objective of the study is to evaluate the effect of GnRH treatment at the time of TET on the conception rate of buffalo embryo recipients. The hypothesis is that treatment with GnRH induces the formation of an accessory corpus luteum (CL), increases the plasma concentrations of progesterone (P4), and increasing conception rate. This study involved 252 Murrah and crossbreeding Murrah x Mediterranea buffaloes,aged from 2 to 15 years, with BCS> 2.5 and belonging to 4 different farms: Fz1 (n=70), Fz2 (n=97), Fz3 (n=63) and Fz4 (n=22). On D0 the recipients received an intravaginal P4 device, associated with i.m. application of 0.5 mg of estradiol benzoate and 0.53 mg of sodium cloprostenol (PGF2α). On D9, the P4 device was removed and 0.53 mg PGF, 400 IU equine chorionic gonadotrophin and 1.0 mg estradiol cypionate were applied i.m. On D18 the buffaloes that had a CL> 10 mm received an embryo and the recipients were divided into two groups: Group GnRH (G-GnRH; n=127) which received 10 μg of acetate of gonadorelin i.m.;and the Control Group (G-CONT; n=125), which did not receive any treatment. A subset of 22 recipients was subjected to Doppler ultrasound examination in the following days: D18, D22, D25, D28 to evaluate the ovulation rate, the formation of an accessory CL and the vascularization of the CLs. After 30 days of TET, a new ultrasound examination was performed to evaluate the conception rate of the recipients. Statistical analyzes were performed using the SAS® GLIMMIX procedure. There was farm effect on conception rate (P <.0001) [Fz1 = 34.2% (24/70); Fz2 = 6.1% (6/97); Fz3 = 31.7% (20/63); Fz4= 31.8% (7/22)]. Lactating buffaloes had a lower rate (12.8%) than the dry buffaloes (31.6%) or heifers (29.4%) [P = 0.004]. No significant difference was observed between the conception rate of the two groups (G-GnRH: 23.6%, G-CONT: 21.6%, P = 0.7). Between the 22 buffaloes examined with Doppler ultrasound, 36% (n = 4/11) of them showed an ovulation following the treatment with GnRH with the subsequent creation of CL accessories. Treatment with GnRH at the time of TET did not increase conception rate in buffaloes.

Lacteos La Delfina - CAPES - Fazenda Ouro Preto - APTA Registro

[1]Mann et al., J Reprod Fertile Suppl., 54: 317-328, 1999 [2]Vecchio et al., Reprod Dom Ani 45,614-618, 2010

82 POSTER PRESENTATIONS Experiences of Artificial Insemination in A Mexican Buffalo Herd 2016 -2018

Jesus BERDUGO1, Luis DE LA CRUZ-CRUZ2, Diego MORALES-CANELA3

1universidad nacional de colombia, Colombia 2Universidad Autonoma Metropolitana, Mexico 3Hacienda Sehualaca, Mexico [email protected], [email protected], [email protected]

The buffalo is a relatively new specie in Mexico with an an exponential growth in the population during the last years. Due to the limitations for the importation of live animals for the reproductive and production improvement programs and the seasonality. Breeders and researchers need to apply different technologies applied in other species, such as artificial insemination. In the Mexican scientific literature, there are no reports of artificial insemination in water buffaloes. To be able to produce animals and milk constantly during the year it is necessary to break the reproductive seasonality. The aim of this paper is to show the experience with artificial insemination, in natural heat and fixed time in a Mexican herd. This work was carried out in the Sehualaca Ranch located in the Municipality of Minatitláan , State of Veracruz, Mexico during 2016 to 2018. The results of the insemination of 275 multiparous adult buffaloes with body condition from 3 to 4.5 (1-5 scale) who underwent a single service, with frozen Murrah or Mediterranean semen. Artifiicial insemination were performed in natural heat detection or after synchronization of ovulation. In the period evaluated, a global pregnancy rate of 60.5% were obtained, corresponding to 165 deliveries, of which 55% (91) and 45% (74) were males and females respectively. 156 inseminations were made at fixed time and 119 with natural estrus, which ended in 94 (60.25%) and 71 (59.66%) births, respectively. Pregnancy rates for Mediterranean and Murrah breeds were 66% and 53% respectively. The effect of the season were observed, being 69% for a favorable breeding season, November-February and 36% for March-May, which is the unfavorable season. It should be noted that outside the reproductive season no pregnancies were obtained from natural estrus because no estrus animals were observed. It is important to note that the pregnancy rates after insemination with synchronization of ovulation are similar to those obtained natural heat, which demonstrate the possibility to apply artificial insemination during all the year.

83 POSTER PRESENTATIONS Histological Characteristics of Buffalo Uterus (bubalus Bubalis) In Folicular and Luteal Phases

Nayara MARQUES, Marlúcia AIRES, Viviane CODOGNOTO, Stella TIRONI, Alessandra SOUZA, Ariana RAMOS, Eunice OBA

School of Veterinary Medicine and Animal Science - UNESP, Department of Animal Reproduction and Veterinary Radiology,Botucatu-SP, Brazil [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

The aim of this study, was characterize the histological aspects of buffalo uterus during its phases aiming to the physiological parameters of the specie. For this experiment accomplishment, blood samples and uterine tissue of 20 crossbred adult buffaloes were collected. Fragments were collected from the regions of the body, middle and upper portions of the right and left sides of uterine horns sides. The samples were fixed in paraformaldehyde and Karnovsky’s solution, infiltrated with resin, cut, stained with hematoxylin and eosin and periodic acid-Schiff, and photographed in a photomicroscope. Uterine horns morphology was analyzed in order to verify the luminal epithelium type, presence and type of uterine glands, glandular epithelium classification and connective tissue constitution. For hormonal evaluation, blood was centrifuged and progesterone dosage was performed by radioimmunoassay technique. The data were analyzed by the least squares method, for analysis of variation, and Tukey test for differences between means with significant result. The animals were classified into the follicular and luteal phases according to morphology and serum progesterone concentration. For the 20 animals used, 8 (38%) were in the follicular phase and 12 (62%) in the luteal phase. The uterus was composed of three layers: mucosa or endometrium, muscular and serous. Endometrial surface of lining epithelium of all the animals was pseudo-stratified to simple cylindrical. Uterine glands were present in large numbers. Few morphological changes were observed in uterine glands. A positive reaction to periodic acid-Schiff was observed in basal and superficial glands of all animals, as well as increased endometrial vascularization in all animals in the follicular phase.

84 POSTER PRESENTATIONS Ultrasonic Doppler as A Tool for Early Pregnancy Diagnosis in Buffalos: Evaluation Of Blood Flow To Corpus Luteum 20 Days After Insemination

Viviane Maria CODOGNOTO1, Felipe Rydygier RUEDIGER1, Paulo Henrique YAMADA1, Ariana DANTAS1, João Carlos Pinheiro FERREIRA1, Nélcio Antonio Tonizza CARVALHO2, Guilherme RIZZOTO3, Andressa Filaz VIEIRA1, Fabiana Ferreira SOUZA1, Eunice OBA1

1São Paulo State University (UNESP), School of Veterinary Medicine and Animal Science, Brazil 2Instituto de Zootecnia (IZ) , Brazil 3Faculty of Veterinary Medicine, University of Calgary, Calgary, Canada, Canada [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

The early pregnancy diagnosis using ultrasonic Doppler (UD) is an important tool to improve reproductive indexes in buffalos, allowing the reduction in the calving interval by detection of non-pregnant animals or early embryonic losses and early utilization re-synchronization protocols and artificial insemination (AI). The objective of this study was to examine 15 multiparous bufallos (Murrah breed) using UD and identify parameters that could be indicators of early pregnancy diagnosis (20 d after AI). The animals were subjected to an estrus synchronization protocol for AI as follows: D0 – Application of P4 intra-vaginal device and estradiol benzoate; D9 – Removal of the device and administration of PGF2α and eCG; D11 – Administration of GnRH analog and performance of AI. Transrectal-DU was performed 20d and repeated at 30d for confirmation of pregnancy. For the exam, a 7.5 MHZ linear transductor was used (esaote, MyLabTMFIVE, Geneva, Italy). The corpus luteum (CL) was evaluated, first using mode B for the diameter (average longitudinal and transversal axis) and posteriorly using colour UD, recording the image for 1min for evaluation of CL irrigation. The irrigation was analyzed through ImageJ (version 1.5 Wayne Rasband National Institute of Health, USA) for the following parameters: luteal area (CLA), irrigated luteal area (ICLA) and irrigated luteal area (%) of the total luteal area (ICLA %). At D20, blood samples were collected for plasmatic P4 concentrations. Results were analyzed through t-tests and qui-square tests (χ2), considering P<0.05. At D20, when comparing pregnant (PG, n=11) vs non-pregnant (NPG, n=4), we observed differences (P<0.05) for the CL diameter, ICLA and ICLA (%) and P4 [ ], at D30 the confirmation of pregnancy was performed with10 out 15 animals being pregnant (93.33% accuracy of 20D). Therefore, the difference between the different parameters studied at D20 between PG and NPG indicates that UD is a tool that can assist on the early pregnancy diagnosis in buffalos, however further studies are still required to improve reliability of the technique.

This research was financially supported through grants from the São Paulo Research Foundation (Grant 2018/13338-3).

85 POSTER PRESENTATIONS Spermal Mobility Of Bubaline Semen After Refrigeration at 5 ° C For 24 Or 48 Hours With Botubov® Extender

Fernando SILVA1, Jaqueline CARVALHO1, João FERREIRA1, Eunice OBA1, Frederico PAPA1, Marc HENRY2

1Unesp FMVZ, Brazil 2UFMG, Brazil [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

Artificial insemination is the biotechnology that has the greatest impact on genetic improvement for agricultural species, and has several advantages such as control of transmission of infectious diseases, zootechnical control and rationalization of reproductive management. In order for gestation after artificial insemination to be effective, the semen must have excellent fertilizer capacity, independent of fresh, refrigerated or frozen use. Artificial insemination is performed predominantly by using frozen semen; however, freezing involves a number of potentially deleterious processes to semen, such as the formation of free radicals and the formation of ice crystals (which does not occur during the cooling process). A new alternative to the use of frozen semen in fixed-time artificial insemination programs (IATF) is the use of refrigerated semen with or without glycerol. The objective was to evaluate the motile values of the refrigerated buffalo semen for up to 48 hours diluted with BotuBov® medium containing cryoprotectant glycerol. Three adult Murrah bulls were used for semen collection using artificial vagina, the semen was diluted in egg yolk medium containing cryoprotectant and evaluated for total motility, progressive motility and rapid spermatozoa in computerized analysis equipment ( CASA) at time 0h (fresh) and after refrigeration at 5 ° C for 24 and 48 hours after collection. The values observed for total motility, progressive motility and fast spermatozoa were: 80.06; 70.43 and 78.73 for fresh semen; 80.63; 60.53 and 77.73 for semen refrigerated for 24 hours; and 74.86; 56.06 and 70.63 for 48-hour refrigerated semen. Therefore, according to the observed, we can conclude that the values of motility were compatible with the parameters indicated by the Brazilian Council of Animal Reproduction.

[1] WATSON, P. F. Animal Reproduction Science., 2000, 60: 481-492. [2] SOUZA, J. D. S.; FERREIRA, W. M. Revista Eletrônica Nutritime, 2007., 4, 456–461.

86 POSTER PRESENTATIONS Response of Bubal Bulls with or Without Conditioning for Semen Collection with Artificial Vagina for Exposure of Estrogenized Females or Not

Fernando SILVA1, Jaqueline CANDIDO2, João FERREIRA2, Eunice OBA2, Frederico PAPA2, Marc HENRY3

1Unesp, Brazil 2Unesp FMVZ, Brazil 3UFMG, Brazil [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

The buffalo bull is probably the easiest species to be conditioned for the collection of semen with the use of an artificial vagina, for this conditioning it is necessary the presence of a female buffalo, there are differences between the animals that are accustomed to make the jump and those which perform only natural coverage. In general, animals that are already conditioned perform the mating regardless of whether or not the female finds estrus. However, those who perform only natural mating need the presence of the female in the estrus. The objective of this study was to verify if for the conditioning the presence of the female in the estrus is necessary To test these affirmations, two groups of animals were studied. Group 1, (n = 3) bulls that were conditioned to perform collection in artificial vagina and group 2 (n = 8) animals that were conditioned to perform only natural mating. To study the response of the animals the following conditions were performed: females in estrus induced with estradiol cypionate (2mg), females that were not in estrus, and pregnant females. Routinely, the artificial vagina is the most commonly used method for collection. The conditioned group performed the samples and allowed the semen collection to be independent of the estrous cycle phase of the female. However, the animals of group 2 performed the mating only on estrogenized females, showing no interest in pregnant females or in other phases of the estrous cycle. The use of estrogenized females is extremely important to initiate the conditioning and the collection of semen using artificial vagina in unconditioned bulls. Bull conditioning avoids the use of more invasive methods such as electroejaculation, and contributes to the advancement of the genetic material distribution of animals from distant locations, in addition to improving animal-man safety.

[1] OHASHI, O.M.; SANTOS, S. S. D.; MIRANDA, M. S. et al. Ver Bras. Reprod. Anim., 2011; 35: 88-94. [2] SANSONE, G.; NASTRI, M. J. F.; FABBROCINI, A. Anim. Reprod. Sci., 2000; 62: 55-76

87 POSTER PRESENTATIONS Effect of The Segregation of Positive Animals on The Incidence Of Tuberculosis In Buffaloes, Corrientes Argentina.

Irina MARTINEZ1, Diana MARTINEZ2, Jesus BERDUGO3, Fabiana CIPOLINI-GALARZA2

1Facultad de Ciencias Veterinarias, UNNE, Argentina 2Facultad de Ciencias Veterinarias, Universidad Nacional del Nordeste, Argentina 3Institulo Latinoamericano para el Estudio del Bufalo, Colombia [email protected], [email protected], [email protected], [email protected]

The buffalo is a bovid that has had great importance in livestock production in Argentina, especially in flooded areas where cattle have low production rates. It has often been inferred that the buffalo is resistant to the vast majority of diseases affecting cattle. However, they are very susceptible to a wide range of infectious agents, among which is Mycobacterium bovis, which is highly pathogenic for buffalo cattle. The bubaline specie is not contemplated in Resolution N ° 128/12 of SENASA that define the Plan for the Control and Eradication of Bovine Tuberculosis in the Argentine Republic. This Plan for cattle determines that any positive reactive animal must be eliminated and sent to slaughter so that epidemiologically the rest of the farms not exposed to the agent. The aim of this paper is to show the effect of animal removal after serological test for tuberculosis disease in two Argentinean herds. In this work, One year old or more buffaloes from two herds E1 and E2 from the province of Corrientes, Argentina, were evaluated. The anocaudal tuberculinization technique were performed, read and evaluated using the same parameters applied to cattle. After the intradermal reaction, positive animals were slaughtered and samples of the lesions compatible with tuberculosis were sent for confirmation by histopathological diagnosis, microbiological culture and PCR. In the E1 during 3 years, once a year they performed the test with the immediate elimination of any positive reaction, so that in 2017 he had an incidence of 1% (1/74), for 2018 it was 0.6% ( 1/152) and in 2019 of 0%. In the E2, the intradermal reaction analysis was performed at higher intervals and not all the positive reactants were eliminated, so in 2014 the incidence was 5% (5/88) in 2016 of 6% (5/81) and finally it remained in 2019 at 6% (5/84). It is concluded that the measure of eliminating positive animals immediately reduces the likelihood that the disease will be maintained and spread within a herd, while the presence of infected animals causes the spread in new animals and the maintenance of the infection.

88 POSTER PRESENTATIONS Vaginal and Rectal Prolapse After Insertion of a Vaginal Devise During Ovulation Synchronization in Buffaloes: Case Report

José KONRAD1, Irina MARTINEZ2, Natalia VALLEJOS2, Alexis BANDEO2, Jesus BERDUGO3, Gustavo CRUDELI2

1Facultad de Ciencias Veterinarias, UNNE. CONICET., Argentina 2Facultad de Ciencias Veterinarias, UNNE, Argentina 3Grupo Biogem, Universidad Nacional de Colombia, Colombia [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

Few reports are in the literature related to genital prolapse, is considered as an emergency that needs immediate attention before any further complication that can lead to a poor prognosis. Vaginal prolapse in buffalo has been reported to occur mostly during the last trimester of gestation or immediately after parturition usually associated with dystocia. However, genital prolapse represents the most common disorder in river buffalo. The aim of this report is to show a very rare case of vaginal and rectal prolapse in an adult non-lactating animal involved in a fixed time artificial insemination program with ovulation synchronized using intravaginal device. The vaginal device was inserted and 1.0 mg of 17b estradiol were injected, three days later the animal start to show signs of discomfort by contracting the pelvic region, showing also sign of pain. First a small portion of the vagina were prolapsed that was manually inserted and are the fourth day the device was expelled and the vagina and rectum prolapsed, with exposure of the mucosa of the two organs. Very shortly the animal was treated surgically and clenbuterol were administrated. Four days later the suture was removed and the animal don’t show more symptoms of prolapse, the animal was discarded from the program. A genetic predisposition to vaginal prolapse have been informed, dietary factors implicated in vaginal prolapse include poor quality forage; hypocalcemia; high estrogenic– content foodstuffs, such as legumes and soybean meal; and overcrowding. Estrogen is a potent inhibitor of bone calcium resorption and elevated levels could plausibly, although not scientifically proved, increase the risk of vaginal prolapse. Individual animal risk factors include obesity, chronic coughing, chronic straining to urinate or defecate. The hypothesis in this case, is that a defect in the placement of the device causes the vagina to feel a foreign body and the contractions begin, which can be increased by the sensitization of the muscles induced by the injected estradiol. Additionally, it must be keep in mind the susceptibility of the animal.

[1] Akhtar MS. Lodhi LA. Ahmad I. Qureshi ZI. Muhammad G. 2012. Serum trace mineral variations in Nili-Ravi buffaloes suffering with prepartum vaginal prolapse in two different agro-ecological zones of Punjab, Pakistan. Theriogenology. 77: 1328-1333. [2] Dhillon KS. Singh BB. Kumar H. Bal MS. Singh J. 2006. Treatment of vaginal prolapse in cows and buffaloes. Vet Rec. 158: 312. [3] Miesner MD. Anderson DE. 2009. Vaginal and uterine prolapse. Food Anim Prac. P: 382-391. [4] Sah SK. Nakao T. 2003. Some characteristics of vaginal prolapse in Nepali buffaloes. J Vet Med Sci. 65: 1213-1215.

89 POSTER PRESENTATIONS Buffalo Meat Economy of India

Amandeep KAUR1, Poonam KATARIA2, Shruti CHOPRA1

1Centre for Agricultural Policy and Market Research,PUNJAB AGRICULTURAL UNIVERSITY, LUDHIANA, India 2Centre for Agricultural Policy and Market Research, PUNJAB AGRICULTURAL UNIVERSITY, LUDHIANA, India [email protected], [email protected], [email protected]

Table1: Major Importers of Indian buffalo meat, 2017-18

Importing Country Value, US$ Million % Share of total

Vietnam 2289.0 56.7

Malaysia 370.0 9.2 Egypt 255.0 6.3 Iraq 156.0 3.9 Saudi Arab 117.0 2.9 Others 849.9 21.0 Total 4036.9 100.0

Animal husbandry has been inseparable from Indian agriculture since times immemorial. India possesses the largest livestock population in the world (512 million heads in 2012), comprising 21.2 per cent buffalo. Keeping into consideration the importance of buffalo in India’s livestock sector, the present study has been conducted to have an insight into buffalo economy of India. The formulations of the study are based on secondary data culled from official sources of India with a time frame of 1990-91 to the latest available. The study brings forth wide inter-state differentials in buffalo population in India. The state of Uttar Pradesh with the highest (28.2 %) buffalo population (19th Livestock Census, 2012) contributes as high as 47.9 per cent to the production of buffalo meat. The three major buffalo meat producing states viz., Uttar Pradesh, Maharashtra and Punjab together contribute two third to the India’s buffalo meat production in 2016-17. Indian buffalo meat has emerged as one of the major agricultural export item, after Basmati Rice. Buffalo meat accounted for 11.5 per cent (worth US$ 3.9 billion) of India’s agricultural exports in 2016-17. Export of buffalo meat has been increasing consistently primarily due to its organic nature and very little domestic demand has fuelled it further. As a buffalo meat producer, India has secured the first position and eighth one as buffalo meat exporter in the global ranking 2016. Vietnam, Malaysia, Egypt, Iraq and Saudi Arabia happened to be the five major export destinations accounting for nearly 79 per cent of the country’s buffalo meat shipments during 2017-18. India has the potential to make inroads into many more markets of developed nations provided policy initiatives are put into place to smoothen the trade flow to the importing countries as per their requirement and norms.

[1] http://apeda.gov.in [2] www.dahd.nic.in [3] www.indiastat.com

90 POSTER PRESENTATIONS Residues of Β-lactam Residues in Milk collected By the Milk Station of The North

Abderraouf GRITLI1, Yahia TEBER2, Rym LAHMAR2, Samir BEN YOUSSEF2

1Directorate General of Military Health, Tunisia 2National School of Veterinary Medicine, Tunisia [email protected], [email protected], [email protected], [email protected]

Antibiotics are important in modern breeding. However, these antibiotics and especially β-lactams can be found as residues in milk from treated cows. The dangers associated with the presence of these residues to public health are toxicological, allergic, microbiological but also technological and environmental.

These risks on public health added to technological problems have led to a gradual strengthening of legislation on veterinary medicine. State took a sum of provisions to fight against these problems, thanks to a regulation, which supervises the veterinary medicine antibiotic from its conception to its use. The state has also put in a gradual development of the dairy sector, measures to prevent and control the risk of antibiotic residues in milk. Plans for monitoring and control of residues in milk are realized every year by the Tunisian State.

Our work is a study on the β-lactam residues in 10 699 milk samples collected between July 2016 and June 2017 in the North West of Tunisia.

The results show contamination by residual β-lactam 0.13% for milk tested. These results highlight the need for the establishment of a systematic monitoring of raw milk produced in Tunisia in order to reduce this rate and reduce health and technological risks as well as economic losses.

91 POSTER PRESENTATIONS The Association of ACAA1 Polymorphisms and Buffalo Milk Production Traits and Gene Function Study

Huaguo HUA, Hui SUN

Huazhong Agricultural University, China [email protected], [email protected]

ACAA1 (acetyl-coenzyme A acyltransferase 1) plays an important role in fatty acids metabolism, which can regulate beta oxidation of fatty acids[1]. Our preliminary studies of buffalo mammary tissue transcriptome sequencing showed that ACAA1 is highly expressed, suggesting ACAA1 may regulate the function of mammary gland.

In this study, we screened the ACAA1 genotypes with 331 Mediterranean buffaloes by MALDI-TOF-MS and its association with buffalo milk production traits. A total of 5 SNPs were identified in ACAA1, which localized at promoter (g.681A>T), introns (g.1868C>T, g.23117C>T, g.24348G>T) and 3′UTR (g.806C>T) regions, respectively. As shown in Figure1, all of the 5 SNPs were found to be significantly associated with buffalo milk production traits (P<0.05).

Moreover, the g.681A>T was located near the promoter active region with strong effects on buffalo milk production traits. Consequently, the potentially promoter activity was tested by Dual-Luciferase Reporter Aassay System. Further functional studies including Electrophoretic Mobility Shift Assay (EMSA) and coimmunoprecipitation (Co-Ip) demonstrated that the A-allele at g.681A>T increased the promoter activity by binding the MyoD transcription factor to improve the transcription of ACAA1 gene.

We further investigated the potential regulation function of ACAA1 in buffalo mammary gland. We found ACAA1 had a higher expression in the middle and late lactation than early lactation, with mainly localized in the cytoplasm and nucleus of buffalo mammary epithelial cells (BMECs). Knockdown of ACAA1 inhibited BMECs proliferation, decreased the secretion of lipid droplet and β-casein, and down-regulated the expression of milk fat and milk protein metabolic pathway related genes (P<0.05). Consistantly, overexpression of ACAA1 increased the cell vitality and cell numbers.

These findings will help in marker-assisted selection for buffalo breeding and the regulatory mechanisms inACAA1 including mammary tissue development and lactation in buffalo.

[1] Wanders R J A, et al. “Peroxisomal fatty acid α- and β-oxidation in humans: enzymology, peroxisomal metabolite transporters and peroxisomal diseases.” Biochemical Society Transactions 29.2(2001):250.

92 POSTER PRESENTATIONS Physiological and Metabolic Responses of Water Buffalo Calves for Effect of Different Weaning Methods

Luis Alberto DE LA CRUZ CRUZ1, Héctor OROZCO-GREGORIO 2, Juan Manuel VARGAS-ROMERO1, José Armando SÁNCHEZ-SALCEDO1, Milagros GONZÁLEZ-HERNÁNDEZ2, Patricia ROLDAN-SANTIAGO3, Jesús Alfredo BERDUGO-GUTIÉRREZ4, Herlinda BONILLA-JAIME1

1Universidad Autónoma Metropolitana, Mexico 2Universidad Autónoma de San Luis Potosí, Mexico 3Universidad del Valle de México, Mexico 4Instituto Latinoamericano del Búfalo, Medellín, Colombia, Colombia [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

The aim of this study is to evaluate and compare the effect of three different weaning methods on physiological and metabolic changes on water buffalo calves. The study included 40 Murrah calves (20 males and 20 females) of 7-8 months of age with an average weight of 229.05 ± 54 Kg. The animals were assigned into four groups of ten animals each: NW= non-weaned calves; AW= weaned abruptly calves and separated permanently from their mothers at day 0; NFW= calves fitted with nose-flaps devices at day -7 before weaning and at day 0 devices were removed and calves were separated permanently from their mothers; and FENW= calves were separated from their mothers through a fence-line at day -7 and at day 0 calves were permanently separated from their mothers. Physiological and metabolic indicators were evaluated on days -7, -6 y -4 before weaning and on days 0, 1, and 3, after the total separation from the mother. All variables were compared with a mixed model for repeated measures, considering the group (NW, AW, NFW, FENW), the time (days), as well as the interaction between group and time as fixed effects, and calf within group as a random effect using the software JMP® 4.0 (SAS Institute Inc.). Statistical differences were considered when p value was < 0.05. Statistical differences were found for effect of group (P< 0.05), time (P< 0.05) and in the interaction group x time (P> 0.0001) in the physiological and energy metabolism variables. FENW calves show higher values of heart rate, respiratory rate, rectal temperature and glucose (P<0.05) during the days -6 and -7 compared with other groups. AW calves show higher values of heart rate, respiratory rate, rectal temperature, glucose and lactate between days 0 and 1 (P<0.05). No statistical differences were found in blood gases and acid-basic, hydric and mineral equilibrium between the groups (P>0.05). The results obtained here show that the abrupt weaning and the separation of the mother through the fence-line produce changes in physiological variables and energy metabolism.

The present study is part of LACC’s Ph.D. program supported by a doctoral fellowship granted by the Consejo Nacional de Ciencia y Tecnología (CONACyT, No. 515743). We thank Mr. Eduardo Maitret and Mr. Manuel Reyes (Agroindustria la Ilama ARIC de R.L de C. V.) for making this study possible by faciliting their buffaloes and expert technical assistance.

93 POSTER PRESENTATIONS Quantification of Methane Emissions: Case of Indian Buffalo

Kashish ARORA, P KATARIA

Punjab Agricultural University, India [email protected], [email protected]

India with only 2.4 % of geographical area is home to 11.5 % of world’s livestock population. India accounts for 56.7 % of world’s buffalo population, 12.5 % cattle, 20.4 % small ruminants and nearly three per cent of poultry population. India’s livestock sector plays an important role in Indian economy as it supports a sizable proportion of small and marginal farm households and provides employment to nearly 9 % of India’s population. India is the largest producer of milk accounting for nearly one fifth of global milk output. India is unique among the major milk producers because more than half of its production is from buffalo, rather than cattle in sharp contrast to only 13 % in the context of world at large. Of late, the concerns are being raised on the environmental effects of livestock sector, which are being fuelled further by increasing global demand for livestock products. It is against this backdrop, the present study aims to ascertain the over time changes in methane emissions by Indian buffalo and the contribution thereof to the total emissions from livestock sector. The formulations of the study are based primarily on secondary data, for four livestock census periods i.e. 1997, 2003, 2007 and 2012, extracted from official data sources of Government of India (GOI). The methane emissions; both enteric and from manure were estimated by using Tier I Methodology given by Inter-governmental Panel on Climate Change (IPCC, 2006). India’s buffalo population, accounting for 21.2 % of livestock population in 2012, contributed as high as 59 per cent of total methane emissions from the livestock sector. The analysis highlighted that methane emission from buffalo population has increased from 5.39 Tg in 1997 to 6.52 Tg in 2012 registering 21 % increase over time. This has been accompanied by decreasing trend in the emissions per unit of milk produced, which should not in any case make the policy planners lose sight of the emergent need to lay thrust on the adaptation and mitigation strategies.

[1] IPCC (2006) IPCC guidelines for national greenhouse gas inventories, prepared by the national greenhouse gas inventories programme (Ed. H. S. Eggleston, L. Buendia, and K. Miwa), IGES, Japan.

94 The Prediction of Lactation Milk Yield from Partial Milk Record in Anatolian Water Buffalo

M.İ. Soysal1, M. Aksel2, E.K. Gürcan1, E. Özkan Ünal1, S. Genç3

1 Namık Kemal University, Faculty of Agriculture, Department of Animal Science, Tekirdag, 2Istanbul Water Buffalo Breeders Association, Istanbul, 3 Ahi Evran University, Faculty of Agriculture, Department of Agricultural Biotechnology, Kırsehir

Abstract This study was researched relationship between total milk yields and partial milk yields of water buffalo reared in Istanbul. The whole data set were obtained as daily milk yield records of water buffalo and fourth lactation number was used in this study. The average lactation period were calculated 216.6 day and the average lactation milk yield were calculated 1360.1 kg. Similarly, the average daily milk yield was calculated 6.26 kg. The relationship between lactation milk yield and additive partial milk yield (30-60-90-120-150-180 day) was analyzed with regression equation. Partial milk yields as independent variables and total lactation milk yield as dependent variable were called and simple and multiple regression analysis methods were used (Y=a+bX and Y=a+b1X1+b2X2). The highest coefficient of determination was identified as the most appropriate model in all models. The correlation coefficient has been found as (r=0.74**) between 30-day partial milk yield and total milk yield and (r=0.88**) between 90-day partial milk yield and total milk yield. Also, the correlation coefficient between total milk yield and 180-day additive milk yield, has been found as (r=0.94**). The relation between total milk yield and additive partial milk yield was analyzed with regression equation. The most suitable regression equation was chosen according to determination coefficient. The determination coefficients (R2) were calculated for 30 and 60 days, using of two independent variables together was 0.77 and using one independent variable (30, 60, 90, 120, 150 and 180 day) partial yields were 0.54, 0.69, 0.77, 0.82, 0.84 and 0.88 respectively. As a result, the partial milk yields could be used to estimate lactation milk yield previously as effectively.

Keywords: Anatolian Water Buffalo, Partial Milk Yield, Lactation Milk Yield

Introduction Water buffalo management was maintained under primitive conditions as general structure in Turkey. Particularly depending on the records of animal, milk and meat yield were kept insufficiently. However, a positive momentum is attained due to the establishment of buffalo breeders unions and improvement projects. Anatolian water buffaloes demonstrated a tendency of quantitative increase last ten years. In 2018 total number of water buffalo, total number of milking water buffalo and total water buffalo milk production were found 180000 heads, 75000 heads and 75000 tons, respectively (Anonymous 2018). Madad et al. (2013), have investigated some factors which influence various yield traits in Azerbaijani water buffalo. In the study, the data was obtained from 2327 first lactation records from 391 buffalo herds, and 1420,54 kg of total milk yield and 99,83 kg of fat yield were found. Moreover, the fat percentage was found to be an average value of 7.01%. Heritability of total milk yield was found as 0,14, on the other hand, fat percentage heritability was found as 35%. Garcia et al. (2013) detected the 244 days average milk yield and lactation length as 864 kg and 240 days, respectively.

Malhado et al. (2013) were found 1546 kg and 252.3 days for milk yield and lactation period respectively. Moreover, according to the study results, they stated that animal selection by using the first lactation could be quick and accurate.

Mohammed A. (2009), was studied in Nili Ravi, Murrah, and Egyptian water buffaloes, the first lactation milk yields were 1854, 1654 and 1185 kg, respectively; the third lactation milk yields were 2396, 2056 and 1678, respectively.

In order to improve milk yield of animal with selection, milk yield capacity of animal could be identified early time of animal. The partial milk yields were used to estimate the total lactation yield.

Total lactation yield was estimated from partial yield with the linear regression equation. This equation was found as (Y=502.919+1.261X), by using 180-days partial yield as (X) and total lactation yield as (Y) for Holstein cattle (Soysal and Küçük 1996). The correlation coefficient between 70-days partial yield and total yield was found as 0.52 in Holstein cattle reared in Italy (Ragnon and Brambilla 1957). Tuncel (1971) was found the coefficient of determination between 30-days and 150-days of partial yield and total yield as 0.596 and 0.861 for Brown Swiss cattle, respectively.

This study was aimed to indicate relationship between total milk yields and partial milk yields of water buffalo reared in Istanbul. The partial milk yields could be used to estimate lactation milk yield previously as effectively.

95 Materials and Methods The animal material of this study was formed by 40 water buffalo reared in Istanbul which was reared in the same management and feeding conditions. In the study, daily milk yield records were used, which were obtained by measuring daily milk yield in the morning and evening from the beginning to the end of lactation period. The whole data set were obtained as daily milk yield records of water buffalo and fourth lactation number was used in this study. The correlation and regression analysis were done between realized lactation milk yield and partial milk yield of the animals, it was attempted to define the partial yield which has the highest coefficient of determination. Partial milk yields weretaken independent variables and lactation milk yield were taken dependent variable. The simple and multiple linear regression analysis methods were used (Y=a+bX) and (Y=a+b1X1+b2X2) in this research. The model with the highest coefficient of determination was determined as the most suitable model (Soysal 2007). Partial milk yields were recorded as 30-60-90- 120-150 and 150-days. Statistical analyses were realized in Statistica software program (Statistica, 1994).

Results The whole data set were obtained as daily milk yield records of water buffalo and fourth lactation number was used in this study. The average lactation period were calculated 216.6 day and the average lactation milk yield were calculated 1360.1 kg. Similarly, the average daily milk yield was calculated 6.26 kg.

The correlation coefficients between values of total milk yield and partial milk yields were presented inTable1. Table 1. The correlation coefficients and significance test results among and between 30-150 days of partial milk yields and total milk yield of animals. Day TMY 30 60 90 120 150 30 0,74** 60 0,83** 0,96** 90 0,88** 0,92** 0,98** 120 0,90** 0,89** 0,96** 0,99** 150 0,92** 0,86** 0,94** 0,98** 0,99** 180 0,94** 0,83** 0,92** 0,96** 0,98** 0,99** **:P<0.01, TMY:Total Milk Yield

The highest coefficient of determination was identified as the most appropriate model in all models. The correlation coefficient has been found as (r=0.74**) between 30-day partial milk yield and total milk yield and (r=0.88**) between 90- day partial milk yield and total milk yield. Also, the correlation coefficient between total milk yield and 180-day additive milk yield, has been found as (r=0.94**).

The relation between total milk yield and additive partial milk yield was analyzed with regression equation. The most suitable regression equation was chosen according to determination coefficient. The regression equations and coefficients of determination were given in Table 2. The determination coefficients (R2) were calculated for 30 and 60 days, using of two independent variables together was 0.77 and using one independent variable (30, 60, 90, 120, 150 and 180 day) partial yields were 0.54, 0.69, 0.77, 0.82, 0.84 and 0.88 respectively.

Table 2. Regression equations and determining coefficients between partial milk yields and total milk yield. Partial Milk Yield Regression Equation (Y=a+bX) Coefficient of Determination (R2) % Partial Milk Yield of 30 day (X) Y=-13+5,748X 54,85 Partial Milk Yield of 60 day (X) Y=-170+3,14X 69,91 Partial Milk Yield of 90 day (X) Y=-187+2,18X 77,66 Partial Milk Yield of 120 day (X) Y=-180+1.70X 82,20 Partial Milk Yield of 150 day (X) Y=-147+1,40X 84,74 Partial Milk Yield of 180 day (X) Y=-72,5+1,18X 88,45 Partial Milk Yield of 30 day (X1) and 60 day (X2)* Y=-80-8,25X1+7.0X2 77,17 Partial Milk Yield of 30 day (X1) , 60 day (X2) and 90 day(X3)* Y=-62-0,89X1-3,72X2+4,88X3 81,79 Not: Y=Predicted milk yield b: Regression coefficient, a: Intercept, *Multiple regression equation (Y=a+b1X1+b2X2)

96 Conclusion In present study, the average lactation period was found 216.6 day and the average lactation milk yield was determined 1360.1 kg. Similarly, the average daily milk yield was found 6.26 kg. In addition, among the animals whose records were taken, the shortest lactation period and the longest lactation period realized as 150 days and 358 days, respectively. Moreover, among recorded animals, it was found that the lowest milk yield was 707,53 kg and the highest milk yield was 2399 kg. The correlation coefficient has been found as (r=0.74**) between 30-day partial milk yield and total milk yield and (r=0.88**) between 90-day partial milk yield and total milk yield. Also, the correlation coefficient between total milk yield and 180-day partial milk yield, has been found as (r=0.94**). The determination coefficients (R2) were calculated for 30 and 60 days, using of two independent variables together was 0.77. As finally, the partial milk yields could be used to estimate lactation milk yield previously as effectively.

References Anonim (2018). TÜİK-Türkiye İstatistik Kurumu, Hayvancılık İstatistikleri. Garcia Y, Fraga LM, Tonhati H, Abreud D, Aspilcueta R, Hernandez A, Padron E, Guzman G, Mora M and Quinonez D (2013). Genetic Parameter Estimates fot Milk Yield and Lactation Length in Buffalo. The 10th World Buffalo Congress and The 7th Assian Buffalo Congress, May 6-8, 2013, Phuket, Thailand. Madad M, Hossein-Zahed NG and Shadparvar AA (2013). A Study of Some Factors Affecting on Productive Traits in Azari Buffaloes of Iran. The 10th World Buffalo Congress and The 7th Assian Buffalo Congress, May 6-8, 2013, Phuket, Thailand Malhado CHM, Ramos AA, Carneiro PLS, Souza JC and Carilo JA (2013). Genetic Parameters for Milk Yield and Lactation Length of Crossbred Buffaloes from Brazil by Bayesian Inference. The 10th World Buffalo Congress and The 7th Assian Buffalo Congress, May 6-8, 2013, Phuket, Thailand. Muhammad A (2009). The Animal of Future . Idara Matbuat-E-Sulemani, Lahore, Pakistan. Ragnon G, Brambılla E (1957). Is it Possiple to Simplify Milk Recording by Limiting the Time of Collecting the Production Data A.B.A. 1957, 26:382, 137 Soysal Mİ, Küçük R (1996). Siyah Alaca Süt Sığırlarında Eklemeli Kısmi Verimler İle Birinci Laktasyon Süt Veriminin Tahmin Olanakları Üzerine Bir Araştırma. Tekirdağ Ziraat Fakültesi Dergisi S.284-293 1996. Soysal Mİ (2007). Biyometrinin Prensipleri, İstatistik I ve II Ders Notları. NKÜ, Ziraat Fakültesi, Yayın No:95, Ders Notu No:64, 237s, Tekirdağ. Statistica, (1994). Statsoft Inc.Tulsaok, Statistica for The Windows TM.Operating System. Tuncel E (1971) Koças Devlet Üretme Çiftliğinde Yetiştirilen Esmer Sığırlarda Kısmi Verimlerden Toplam Süt Veriminin Tahmini ve Bundan Seleksiyonda Yararlanma Olanakları Üzerine Araştırmalar Doktora Tezi, Ankara Üniversitesi Ziraat Fakültesi, Zootekni Bölümü 97 The Comparison of Lactation Curve for Anatolian Water Buffalo Reared in Istanbul

M.İ. Soysal1, M. Aksel2, E.K. Gürcan1, E. Özkan Ünal1, S. Genç3

1 Tekirdağ Namık Kemal University, Faculty of Agriculture, Department of Animal Science, Tekirdağ. 2Istanbul Water Buffalo Breeders Association, Istanbul. 3 Kırşehir Ahi Evran University, Faculty of Agriculture, Department of Agricultural Biotechnology, Kırşehir.

Abstract Biometry of lactation was investigated with different mathematical models for Anatolian water buffalo reared in Istanbul. The data set were obtained as daily milk yield records of water buffalo in present study. The animal material was originated from Italy and Turkey. All animals were grouped for lactation number. Wood, quadratic, Wilmink, logarithmic quadratic models were selected from within the models. The average lactation period were calculated 228.21 and 243.39 day and this different were not found very important as statistically for animals which were originated from Turkey and Italy respectively. The average lactation milk yield were calculated 1309.1 and 1747.8 kg and the average daily milk yield were calculated 5.65 and 7.17 kg and this different were found very important as statistically (P<0.01) for animals that were originated from Turkey and Italy respectively. Determination coefficients (R2) were calculated for Wood, quadratic, Wilmink and logarithmic quadratic models as 0.71-0.70, 0.66-0.64, 0.69-0.69 and 0.72-0.70 respectively for animals which were originated from Turkey and Italy. At the same time, persistency (S), maximum milk yields (Ymax) and the time of obtaining the milk yield (Tmax) values were calculated for Wood model. These values were found as persistency (S), maximum milk yield (Ymax) and the time of obtaining the milk yield (Tmax) are 6.03 and 6.19, Ymax values are 7.87and 10.35 kg and Tmax values are 31.05 and 35.20 day respectively for animals which were originated from Turkey and Italy. In this study, daily milk yield records of Anatolian water buffaloes were used in modeling lactation curves with four models and compares origined of animal and lactation number.

Keywords: Anatolian Water Buffalo, Lactation Curve, Wood Model, Wilmink Model, Biometry of Lactation

Introduction In spite of the fact that water buffalo breeding in Turkey, has decreased until 2012 but after this year water buffalo number has started to increase again. Unfortunately, it is still generally done under primitive conditions. The number of water buffaloes in the Asian is shows a tendency to increase. in Turkey, water buffalo numbers decrease rapidly by reason of information and execution of wrong policies until 2010. In recent years, owing to the right decisions, positive executions have been taken towards this issue. Considering statistical data, by the end of 2018, the total number of water buffalo has increased to 180000 heads, the number of milking buffalo has increased to 75000 heads and the amount of buffalo milk production has reached to 76000 tons (Anonymous 2018). Water buffalo’s milk is used kaymak, yogurt, cheese, and ice- cream in production. From another hand, water buffalo meat is used in sucuk production (Soysal 2009).

Mohammed A. (2009), noted that Nili Ravi buffaloes in Pakistan, had a mean lactation period of 317 days and a mean lactation milk yield of 2219 kg. Additionally, the lactation duration and milk yield of swamp buffaloes, which were raised in Philippines, have been reported as 187,3 days and 1481 kg, respectively. Furthermore, it has been reported that, in Nili Ravi, Murrah, and Egyptian buffaloes, the first lactation milk yields were 1854, 1654 and 1185, respectively, while the third lactation milk yields were 2396, 2056 and 1678. The same researcher stated that Italian buffaloes produce 2587 kg of milk on average 322,9 days.

A study about the biometrics of the lactation curve in Iranian buffalo, in which some environmental factors (year of birth, year of calving, calving age, calving season) and traits of various lactation curves (starting milk yield, peak milk yield, the slope of the rise and fall of lactation curve when the peak yield occurs, persistence and 240-day milk yield) were researched, has been done. In the study, calving age, calving season, year of birth and year of calving were found to be statistically important factors on the lactation curve characteristics (P <0.05). In addition to this, it was reported that non- genetic factors have a significant role in determination of total milk yield and lactation in buffaloes (Hasanpur et al. 2013). Garcia et al. (2013), have worked on genetic parameter estimations of buffalo milk yield and lactation duration and used the data consisted of 2575 lactation records from 1377 buffaloes. The study indicated that 244-day mean milk yield and lactation period length were 864 kg and 240 days, respectively. Additionally, while the heritability levels of milk yield and lactation length were detected to be 0.15 and 0.13, respectively, the genetic correlation between these properties were calculated as 0,63.

Shape of lactation curve is a significant criteria besides milk yield. Accordingly, regarding to the shape of lactation curve, it is possible to obtain some conclusions on lactation milk yield of animal. For instance, a cow showing little change during lactation is more preferred than a cow giving a large part of milk at the beginning and a little amount after peak yield. In this case, the first animal’s curve is called flat lactation curve, and the second animal’s curve is called the steep lactation curve. In animal breeding practice, animals with flat lactation curve are preferred over the ones with steep lactation curve (Akbulut et al. 1994).

98 Şahin et al. (2013), have compared eight different lactation curve models by using milk yield records of Anatolian water buffaloes which bred in different farming conditions in 2011-2013. In this study, Wood, Cobby and Le Du, exponential, parabolic exponential, inverse polynomial, quadratic, logarithmic quadratic and logarithmic linear models were used in defining lactation curve. Determination coefficient (R2) and residual standard deviation (RSD) coefficient were employed as a criterion to identify the best model that defines lactation curve. The results indicated that quadratic logarithmic and quadratic models that produce the highest R2 and the lowest RSD values are showing the best fit.

In the study, daily milk yield records of Anatolian water buffaloes were used in modeling lactation curves and determining the most suitable model for originate in Turkey and Italy.

Material and Method This study was realized with different mathematical models for Anatolian water buffalo reared in Istanbul. The data set were obtained as daily milk yield records of water buffalo in present study. The animal material was originated from Italy and Turkey. Total 94 head animal were used in this study. All animals were grouped for lactation number. Wood, quadratic, Wilmink, logarithmic quadratic models were selected from within the models. At the same time, persistency (S), maximum milk yields (Ymax) and the time of obtaining the milk yield (Tmax) values were calculated for Wood model. In the study, persistance (S) values were found to be S=-(1+b).lnc. Moreover, the day that the highest milk yield was obtained (Tmax) and maximum milk yield (Ymax), were calculated to be Tmax=b/c, Ymax=a.(b/c)b.e-b respectively (Wood 1967). The function formulas of the models were given in Table 1. Modeling processes and parameter estimates of the models used were made in statistical package software Statistica (Statistica, 1994).

Table 1. The models used in this study Model Function Wood Y(t)=atb e-ct Quadratic Y=a+bx+cx2 Wilmink Y(t)=a + be-kt + ct Logarithmic Quadratic Y=β0 + β1 T + β2 T2 + β3 loge(T2)

Findings and Discussion In the study, daily milk yield records of 94 heads Anatolian water buffalo have been kept to be used in modeling lactation curve.

The average lactation period were calculated 228.21 and 243.39 day and the average lactation milk yield were calculated 1309.1 and 1747.8 kg and this different were found very important as statistically (P<0.01) for animals which were originated from Turkey and Italy respectively. Similarly, the average daily milk yield were calculated 5.65 and 7.17 kg and this different were found very important as statistically (P<0.01) for animals which were originated from Turkey and Italy respectively. In Table 2, 3 and 4 lactation period, lactation milk yield and daily avareage of milk yield values of animals are presented according to origin and lactation number.

Table 2. Lactation period, lactation milk yield and daily average of milk yield values of animals according to origin. Characteristics Origin N Mean S P Lactation period Turkey 43 228,21 50,74 0,18 Italy 51 243,39 57,10 Lactation milk yield Turkey 43 1309,1A 440,6 0,01 Italy 51 1747,8B 614,8 Daily average of milk yield Turkey 43 5,65A 1,14 0,01 Italy 51 7,17B 1,71

Table 3. Lactation period, lactation milk yield and daily average of milk yield values of animals according to lactation number for Turkey origin

Characteristics Lactation Number N Mean S P Lactation period 2 4 216,8 54,1 0,52 3 19 238,2 47,9 4 20 221,1 53,6 Lactation milk yield 2 4 972 374 0,08 3 19 1456,9 433,8 4 20 1236,2 421,3 Daily average of milk yield 2 4 4,42B 0,76 0,02 3 19 6,05A 1,05 4 20 5,52AB 1,12

99 Table 4. Lactation period, lactation milk yield and daily average of milk yield values of animals according to lactation number for Italy origin

Characteristics Lactation Number N Mean S P

Lactation period 2 6 283,7A 60,3 0,01 3 25 258,8A 61,8 4 20 212,1B 30,7 Lactation milk yield 2 6 1800AB 428 0,03 3 25 1946A 701 4 20 1486B 448 Daily average of milk yield 2 6 6,68 1,95 0,54 3 25 7,42 1,48 4 20 7,00 1,94

Determination coefficients (R2) were calculated for Wood, quadratic, Wilmink and logarithmic quadratic models as 0.71- 0.70, 0.66-0.64, 0.69-0.69 and 0.72-0.70 respectively for animals which were originated from Turkey and Italy and results were given in Table 5

Subsequently, determination coefficients were calculated between 0,66 and 0,72 but these coefficients werenot assessment as very highest.

Table 5. The determination coefficient (R2) of models for general groups. Models Origin N Determination Coefficient (R2) % Wood Turkey 43 71,01 Italy 47 70,14 Quadratic Turkey 43 66,67 Italy 47 64,46 Wilmink Turkey 43 69,89 Italy 47 69,13 Logarithmic Quadratic Turkey 43 72,00 Italy 47 70,78

The parameter estimations related to these models were demonstrated in Table 6. According to the results, initial milk yields for Wood model was detected to second lactation 4,31-5,99 kg, thirty lactation 5,03-6,35 kg and fourth lactation 4,58-5,71 kg respectively for animals which were originated from Turkey and Italy. All models were shown as detailed for origin and lactation number in Table 7.

Table 6. Parameter estimations of all models according to origin of water buffalo. Models Origin n a b c d R2 % SSE Wood Turkey 43 4,75B 0,22 0,007 - 71,01 220,3 Italy 47 6,03A 0,22 0,006 - 70,14 401,4 Quadratic Turkey 43 7,77B -0,005 -0,00006 - 66,67 268,4B Italy 47 9,92A -0,008 -0,00007 - 64,46 490,3A Wilmink Turkey 43 9,47B -3,44 -0,02 - 69,89 232,6B Italy 47 11,95A -4,41 -0,03 - 69,13 443,7A Logarithmic Quadratic Turkey 43 4,24B -0,05 0,00004 0,80 72,46 202,5B Italy 47 5,28A -0,07 0,00007 1,02 70,78 370,1A R2: determination coefficient, SSE: sum of square error

100 Table 7. All models according to origin and lactation number.

Models Origin Lactation Number Equations Wood Turkey 2 Y=4,31.t0,20.2,71-0,007.t 3 Y=5,03.t0,20.2,71-0,006.t 4 Y=4,58.t0,25.2,71-0,008.t

Italy 2 Y=5,99.t0,19.2,71-0,005.t 3 Y=6,35.t0,24.2,71-0,007.t 4 Y=5,71.t0,20.2,71-0,006.t Quadratic Turkey 2 Y=6,36-0,01.t+0,000005.t2 3 Y=8,10-0,002.t-0,00006.t2 4 Y=7,75-0,005.t-0,00007.t2

Italy 2 Y=9,57-0,014.t-0,00003.t2 3 Y=11,04-0,017.t-0,00005.t2 4 Y=8,62+0,005.t-0,0001.t2 Wilmink Turkey 2 Y=7,24-2,29.2,71-0,05.t-0,01.t 3 Y=9,89-3,63.2,71-0,05.t-0,02.t 4 Y=9,52-3,48.2,71-0,05.t-0,03.t Italy 2 Y=10,94-3,43.2,71-0,05.t-0,02.t 3 Y=13,08-4,90.2,71-0,05.t-0,03.t 4 Y=10,95-4,13.2,71-0,05.t-0,02.t Logarithmic Quadratic Turkey 2 Y=4,15-0,05.t+0,00003.t2+0,59.log(t2) 3 Y=4,51-0,05.t+0,00004.t2+0,80.log(t2) 4 Y=4,10-0,06.t+0,00005.t2+0,85.log(t2)

Italy 2 Y=5,08-0,06.t+0,00008.t2+0,94.log(t2) 3 Y=5,34-0,08.t+0,0001.t2+1,23.log(t2) 4 Y=5,26-0,04.t+0,00001.t2+0,78.log(t2)

At the same time, persistency (S), maximum milk yields (Ymax) and the time of obtaining the milk yield (Tmax) values were calculated for Wood model and presented in Table 8. These values were found as persistency (S), maximum milk yield (Ymax) and the time of obtaining the milk yield (Tmax) are 6.03 and 6.19, Ymax values are 7.87and 10.35 kg and Tmax values are 31.05 and 35.20 day respectively for animals which were originated from Turkey and Italy.

Table 8. Persistance, Ymax and Tmax values of Wood model according to lactation number and origin of animal. Models Origin Lactation Number S Ymax Tmax

Wood Turkey 2 5,95 6,90 28,57 3 6,13 8,30 33,33 4 6,03 8,43 31,25

Italy 2 6,30 9,89 38 3 6,15 11,67 34,28 4 6,13 9,43 33,33

The observed and predicted yield with respect to time is calculated by equation of models. The observed and predicted yield were charting as a line graph. The most fitted line graphs were selected with regard to model. These graphs were grouped according to origin and lactation number and were given in Figure 1-6.

101 Figure 1. The lactation curves of Wood and quadratic models for second lactation and originated from Turkey (Yield- Month).

Figure 2. The lactation curves of Wood and quadratic models for third lactation and originated from Turkey (Yield-Month).

Figure 3. The lactation curves of Wood and quadratic models for fourth lactation and originated from Turkey (Yield-Month).

Figure 4. The lactation curves of Wood and quadratic models for second lactation and originated from Italy (Yield-Month).

Figure 5. The lactation curves of Wood and quadratic models for third lactation and originated from Italy (Yield-Month).

102

Figure 6. The lactation curves of Wood models for third lactation and originated from Italy (Yield-Month).

Some studies were summarized such as in a study conducted on Iranian buffaloes, biometry of lactation curve was researched. In the framework of this study, increase and decrease slope of lactation curve, persistence, and 240-day milk were researched. Thereby, it was found that calving age, calving season, year of birth, and calving year were statistically important factors for the characteristics of lactation curve (P<0.05) (Hasanpur et al. 2013).

Garcia et al. (2013) have found 244-day mean milk yield and lactation length as 864 kg and 240 days, respectively. From another hand, heritability of milk yield and lactation length have been indicated to be 0.15 and 0.13, respectively, while the genetic correlation between these properties, has been calculated as 0.63. Malhado et al. (2013) was found milk yield and lactation period of hybrid buffaloes as 1546 kg and 252.3 days, respectively.

Şahin et al. (2013), were compared 8 different lactation curve models by using control day milk yield records of Anatolian water buffaloes. In order to define lactation curve, Wood, Cobby and Le Du, Exponential, Parabolic Exponential, Inverse Polynomial, Quadratic, Logarithmic Quadratic and Logarithmic Linear models were used in the conduct of this study. Subsequently, it is detected that Quadratic Logarithmic and Quadratic models that produce the highest R2 and the lowest RSD values, are showing the best fit.

Gürcan et al. (2011) have used quadratic logarithmic, linear, logarithmic quadratic, linear hyperbolic, inverse polynomial and Wilmink models. Subsequently, among the models used in the research, the most appropriate and the highest adjusted determination coefficient value was found in logarithmic quadratic model (0.97). On the other hand, adjusted determination coefficient value was detected as 0.86 for the Wilmink model. Furthermore, according to the results, it was stated that 77% of the animals have showed typical lactation curve, while 23% have showed an atypical lactation curve. For present study were not found any atypical lactation curve for all lactation.

In this study, Wood, quadratic, Wilmink and logarithmic quadratic models were used. However, among the models used the coefficient of determination was found to be approximately similar for the all models.

The average lactation period were calculated 228.21 and 243.39 day, the average lactation milk yield were calculated 1309.1 and 1747.8 kg and the average daily milk yield were calculated 5.65 and 7.17 kg and this different were found very important as statistically (P<0.01) for animals that were originated from Turkey and Italy respectively.

Result In this study, at the same time performance of animals were comprised for lactation milk yield and daily milk yield which is originated from Turkey and Italy and animals that were originated from Italy were detected high performance from originated from Turkey (P<0,01). Therefore these animals could be suggestible as parents of new generation. In the study conducted with the daily milk yield records, Wood, quadratic, Wilmink and logarithmic quadratic models were used and Wood and quadratic model were found most suitable model from the other models according to determination coefficient with observed and predicted values. Wood model is preferable from quadratic model because it could be use to calculate some parameters as persistency (S), maximum milk yield (Ymax) and the time of obtaining the milk yield (Tmax) in addition.

103 References Anonim (2018). TÜİK-Türkiye İstatistik Kurumu, Hayvancılık İstatistikleri. Akbulut Ö, Emsen H, (1994).Atatürk Üniversitesi Tarım İşletmesinde Yetiştirilen Esmer, İleri Kan Dereceli Esmer Melezleri ile Siyah Alaca Sığırların Süt Verim Özellikleri ve Laktasyon Eğrisi Parametrelerine Etkili Faktörler. Atatürk Üniversitesi Ziraat Fakültesi Dergisi, 25 (3), 327-343. Garcia Y, Fraga LM, Tonhati H, Abreud D, Aspilcueta R, Hernandez A, Padron E, Guzman G, Mora M and Quinonez D (2013). Genetic Parameter Estimates for Milk Yield and Lactation Length in Buffalo. The 10th World Buffalo Congress and The 7th Assian Buffalo Congress, May 6-8, 2013, Phuket, Thailand Gürcan EK, Soysal Mİ, Küçükkebapçı M, Yüksel MA ve Genç S (2011). Mandaların laktasyon eğrisinin farklı modellerle karşılaştırılması. 7.Ulusal Zootekni Bilim Kongresi-Adana,2011. Hasanpur K, Aslaminejad AA, Kıanzad D, Naderfard HR, Seyyedalian SAR and Javanmard A (2013). The Study of Individual Lactation Curves of Two Iranian Buffalo Ecotypes. The 10th World Buffalo Congress and The 7th Assian Buffalo Congress, May 6-8, 2013, Phuket, Thailand Şahin A, Ulutaş Z, Yıldırım A, Yüksel A, Genç S. (2014). Anadolu Mandalarında Farklı Laktasyon Eğrisi Modellerinin Karşılaştırılması. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 20 (6): 847-855. Malhado CHM, Ramos AA, Carneiro PLS, Souza JC andCarilo JA (2013). Genetic Parameters for Milk Yield and Lactation Length of Crossbred Buffaloes from Brazil by Bayesian Inference. The 10th World Buffalo Congress and The 7th Assian Buffalo Congress, May 6-8, 2013, Phuket, Thailand. Mohammed A (2009). The Animal of Future. Idara matbuat-E-Sulemani, Lahore, Pakistan. Soysal MI (2009). Manda ve ürünlerinin üretimi. Tekirdağ, Yayın no: 978-9944-5405-3-7, Sayfa:161-171, Tekirdağ. Statistica, (1994). Statsoft Inc.Tulsaok, Statistica for The Windows TM. Operating System. Wood, P.D.P. (1967). Algebraic Model of LactationCurve in Cattle. Nature, 216, 164-1.

104 Cooling Perfomance Assessment of Fogging System in Anatolian Water Buffalo with Regard to Animal Welfare*

M. Ihsan Soysal1 Israfil. Kocaman2* H. Comert. Kurc2

1Namik Kemal University Agricultural Faculty Animal Science, TR-59030 Tekirdag Turkey. 2Namik Kemal University Agricultural Faculty Biosystems Engineering Department, TR-59030 Tekirdag Turkey. *[email protected]

Abstract This research was carried out to evaluate the cooling performance of fogging system which were installed in the water buffalo barn with regard to animal welfare. Two experimental groups which have ten water buffaloes for each group were formed based on genetic similarities, lactation numbers and birthdates in the studied farm. The first group was housed in the Barn-I where the indoor temperature was controlled and the fogging system was installed, and the second group was housed in the Barn-II under existing farmer conditions. Same feeding ration was implemented for two groups. The cooling efficiency values of the fogging system installed in the Barn-I were varied between 45.6% and 85.7%. Temperature- Humidity Index values were 19.9 and 23.1 in the Barn-I and 19.5 to 26.4 in the Barn-II. As a result, a well-planned fogging system for animal welfare will be extremely useful in eliminating or reducing the negative effects of temperature stress. Keywords: Water buffalo, fogging system, animal welfare, cooling performance

*The study was supported by TOVAG of TUBITAK with project number 115O602.

105 Cooling Perfomance Assessment of Fogging System in Anatolian Water Buffalo with Regard to Animal Welfare*

M. Ihsan Soysal1 Israfil. Kocaman2a H. Comert. Kurc2

Abstract This research was carried out to evaluate the cooling performance of fogging system which were installed in the water buffalo barn with regard to animal welfare. Two experimental groups which have ten water buffaloes for each group were formed based on genetic similarities, lactation numbers and birthdates in the studied farm. The first group was housed in the Barn-I where the indoor temperature was controlled and the fogging system was installed, and the second group was housed in the Barn-II under existing farmer conditions. Same feeding ration was implemented for two groups. The cooling efficiency values of the fogging system installed in the Barn-I were varied between 45.6% and 85.7%. Temperature- Humidity Index values were 19.9 and 23.1 in the Barn-I and 19.5 to 26.4 in the Barn-II. As a result, a well-planned fogging system for animal welfare will be extremely useful in eliminating or reducing the negative effects of heat stress. Keywords: Water buffalo, fogging system, animal welfare, cooling performance

*The study was supported by TOVAG of TUBITAK with project number 115O602.

Introduction

The thermal stress is the one of most important environmental factor in terms of animal production and welfare. Its effects on animals and their performance have been well defined. As rectal temperature increases with high indoor temperature, feed consumption and milk yield decrease. When the indoor temperature rises beyond the normal limits (5-25 ° C), the relative humidity also increases. It adversely affects the animal’s thermoregulation ability. The high yielding animals are more affected than low yielding animals because they are metabolically more active and they store more extra heat. Some measurements such as fogging, showering, and fogging with fans should be taken in order tolerate thermal stress of farm animals (Gorgulu, 2013).

The indoor temperature for the water buffalo is more important than the other cattle. The water buffalo is a group of homoeothermic animals, like the other cattle, and their skin contains sweat gland less than other cattle (Borghese, 2005). In addition, the skin of the water buffalo is thicker than the skin of cattle and their body hair density is much lower than other cattle. While this value is 100-200/cm2 for water buffalo, it is 1000/cm2 for other cattle. This situation limits heat dissipation and evaporation with perspiring and feed consumption, growth rate, milk production and fertility are negatively affected. (Thomas, 2004).

In the traditional farms, water buffaloes cool off in accumulated waste water without hygienic condition (Figure 1). Some farms go to the way of construction of the refreshment ponds. Also, this way has some problems in terms of environment and animal health.

Figure 1. Accumulated waste water using by water buffaloes

106 Material and Method

This study was conducted in a water buffalo farm which had sufficient flock capacity. This farm is located in Thrace Part of Istanbul Province and a member of Water Buffalo Breeder Association. The study area is located at 41° 12 ‘northern latitude and 28° 44’ east longitude and its average altitude from sea level is 119 m. According to meteorological records, the average annual temperature is 13.8 ºC and the average annual relative humidity is 84.5% for research area (Anonymous, 2019).

Two experimental groups which had ten water buffaloes for each group were formed based on genetic similarities, lactations numbers and birthdates in the investigated farm. Same feeding program was implemented for each group. The first group was housed in the Barn-I where the indoor temperature was controlled and the fogging system was installed, and the second group was housed in the Barn-II under existing farmer conditions. The ambient temperature was kept below 25 ° C, which is considered to be the initial point of the temperature stress in the Barn-I.

In the fogging system; the fresh water which was passed through the filter system was directed to the lines with a pressure of 70-120 bar by high-pressure electric water pump. Water particles were fogged lower than 20 µm to indoor environment by 0.5 mm diameter nozzles. The fogging lines were placed above 1.5 m from the withers height of water buffalo in the roof space and at the center of the standing platform. The fogging system was shown in the Figure 2.

Figure 2. The fogging system The indoor temperature and relative humidity values were continuously measured at 10 minutes intervals using by the thermo-hygrometer for 24 hours along a year. Surface body temperature values of water buffaloes were measured by using infrared thermometer in order to monitor evaluation of fogging system. Also, rectal temperature of water buffaloes were recorded by thermometer.

The cooling efficiency of the fogging system and the temperature humidity index were calculated (Marai et al. 2009, Mutaf, 2012).

107 Results and Discussion In this study, average indoor and outdoor temperature values, average body surface and rectal temperature values of water buffaloes were recorded. The obtained data in Barn-I and Barn-II were given in Table 1 and Table 2, respectively for summer season.

108 109 Barn- I and Barn-II were compared each other in terms of average body surface and rectal temperatures of water buffaloes (Figure 3).

Figure 3.The body surface and rectal temperature changes of water buffaloes in the Barn-I and Barn-II for summer season. As it has been seen in Figure 3 that the values of body surface and rectal temperatures in the Barn I was lower than in Barn-II.

In this study, cooling efficiency of the fogging system were calculated. The values were given in Table 3.

Table 3 has showed that the cooling efficiency of fogging system varied between 45.6% and 85.7%. The cooling efficiency increases as the difference between dry and wet bulb temperatures increase. If the percentage values of the cooling efficiency increase from zero, the performance of fogging system increases (Mutaf, 2012). Temperature-Humidity Index values were given in Table 4 for Barn-I and Table 5 for Barn-II.

110 While Temperature-Humidity Index values for Barn-I varied between 19.9-23.1, 11.9-20.5 and 15.9-21.8, these values differed between 19.5-26.4, 9.6-21.6 and 14.8-23.6 for Barn-II based on daily maximum average, daily minimum average and monthly average data, respectively. It showed that the temperature stress was very serious level in summer season based on daily maximum average data for Barn-II. Based on monthly average data, it was also serious level for summer season.

Conclusion The cooling efficiency values of this system varied between 45.6% and 85.7%. Temperature-Humidity Index values ranged from 15.9 to 21.8 in the Barn-I and from 14.8 to 23.6 in the Barn-II. It was found that the fogging system had great impact to prevent thermal stress in the hot summer months

References Anonymous (2019). ‘‘Meteorological Data in Catalca District of Istanbul Province”. Metorology General Directorate, Ankara, http://www.mgm.gov.tr. Borghese. A. 2005. ‘‘Buffalo Production and Research’’, FAO Reu. Tech. Ser., 67. 216-220. Gorgulu, M. (2013). Heat stress and feeding, http://www.muratgorgulu.com.tr, Last access date: 06 March 2018. Marai, I.F.M., Daader A.H., Soliman A.M., El-Menshawy S.M.S. 2009.’‘Non-Genetic Factors Affecting Growth and Reproduction Traits of Buffaloes under Dry Management housing in Egypt’’ Livestock Research for Rural Development 21(3),1-12. Mutaf, S. (2012). “The Climatic Environment and Detection Principles of Animal Barns with Engineering Approach” (First Edition). Ankara: Publications of Ministry of Food, Agriculture and Livestock. Thomas, C.S. (2004). Milking Management of Dairy Buffaloes. Doctoral thesis, Swedish University of Agricultural Sciences, Uppsala. Acta Universitatis Agriculturae Sueciae, Agraria, 455.

111 Organized By

Supported By

Organization Office E-mail : [email protected] Phn : +90 312 441 0700 www.arber.com.tr