Editors Wisitiporn Suksombat Mariena Ketudat-Cairns Kanchana Punyawai
Rangsun Parnpai Kanokwan Srirattana Arthip Limcharoen
Congresses Secretariat Embryo Technology and Stem Cell Research Center School of Biotechnology Institute of Agricultural Technology Suranaree University of Technology E-mail: info@wbc2013, www.wbc2013.com
Editorial Board Buffalo Reproduction Symposium 1. Prof. Yindee Kitiyanant 2. Prof. Dr Kehuan Lu 3. Prof. Dr. Dheer Singh 4. Dr. Sunpetch Sophon 5. Assoc. Prof. Petai Pongpiachan 6. Assoc. Prof. Dr. Yupaporn Chaiseha 7. Assoc. Prof. Dr. Bunlue Kornmatitsuk 8. Assoc. Prof. Dr. Suneerat Aiumlamai 9. Assoc. Prof. Dr. Siriwat Suadsong 10. Asst. Prof. Pongthorn Suwannatada 11. Asst. Prof. Dr. Theerawat Tharasanit 12. Dr. Takashi Nagai 13. Dr. Kei Imai 14. Dr. Tamas Somfai 15. Dr. Giorgio Antonio Presicce 16. Dr. Vibuntita Chankitisakul 17. Dr. Nutthee Amin 18. Dr. Anucha Sathanawongs 19. Dr. Krittiya Lertchunhakiat 20. Dr. Thuchadaporn Chaikhun 21. Mr. Anawat Sangmalee 22. Ms. Jakkhaphan Chasombat 23. Mr. Ashit Pual
Buffalo Genetics and Breeding Symposium 1. Prof. Dr. Kanchana Markvichitr 2. Prof. Masroor Ellahi Babar 3. Prof. Leopoldo Iannuzzi 4. Assoc. Prof. Dr. Monchai Duangjinda 5. Assoc. Prof. Dr. Marina Ketudat-Cairns 6. Assoc. Prof. Dr. Neramit Sukmanee 7. Asst. Prof. Dr. Sirinporn Sindhuvanich 8. Asst. Prof. Dr. Skorn Koonawootrittririon 9. Asst. Prof. Dr. China Supakorn 10. Asst. Prof. Dr. Amornrat Molee 11. Asst. Prof. Dr. Wuttigrai Boonkum 12. Dr. Nalinee Imboonta 13. Dr. Supawadee Manatrinon 14. Dr. Warangkana Kitpipit 15. Dr. Yuanyuan Liang 16. MS. Kanokwan Srirattana 17. Mr. Phakphume Saowaphak
Buffalo Nutrition and Feeding Symposium 1. Assoc. Prof. Dr. Wisitiporn Suksombat
Buffalo Health Symposium 1. Prof. Dr. Chaleow Salakij 2. Assoc. Prof. Dr. Anudep Rungsipipat 3. Assoc. Prof. Dr. Wijit Banlunara 4. Asst. Prof. Dr. Somporn Techangamsuwan 5. Asst. Prof. Dr. Burin Nimsuphan 6. Asst. Prof. Khamphee Pattanatanang
7. Asst. Prof. Dr. Sonthaya Tiawsirisup 8. Asst. Prof. Dr. Nareerat Viseshakul 9. Asst. Prof. Dr. Worakij Cherdchutham 10. Asst. Prof. Dr. Sirikachorn Tangkawattana 11. Asst. Prof. Dr. Piyanan Taweethavonsawat 12. Dr. Kidsadagon Pringproa
Buffalo Physiology Symposium 1. Assoc. Prof. Dr. Apassara Choothesa 2. Assoc. Prof. Dr. Jatuporn Kajaysri 3. Assoc. Prof. Dr. Witaya Suriyasathaporn 4. Asst. Prof. Dr. Siriwan Prapong 5. Dr. Chaiwat Boonkaewwan 6. Dr. Narudee Kashemsant
Buffalo Production and Management Symposium 1. Prof. Dr. Ibrahim Soliman
Buffalo Socio-Economic and Sustainable Production Symposium 1. Assoc. Prof. Dr. Kwunkamol Donkwa 2. Asst. Prof. Dr.Kanchana Sukanthasirikul 3. Asst. Prof. Chanisa Maneeratrungrord 4. Mrs. Ancharlie Na Chiangmai 5. Dr. Kampanat Vijitsrikamol 6. Dr. Worapote Suthisai 7. Dr. Boonchauy Boonme 8. Dr. Kulapa Kuldilok
Buffalo Meat and Meat Products Symposium 1. Professor Dr. Sanchai Jaturasitha 2. Assoc. Prof. Dr. Suthipong Uriyapongson 3. Asst. Prof. Dr. Sujate Chaunchom 4. Asst. Prof. Dr. Chaiyawan Wattanachant
Buffalo Milk and Milk Products Symposium 1. Assoc. Prof. Dr. Somkiert Prasanpanich 2. Asst. Prof. Dr. Manote Sutheerawattananonda 3. Asst. Prof. Dr. Piyawan Gasaluck 4. Asst. Prof. Dr. Surasak Kochapakdee 5. Asst. Prof. Dr. Pipat Lounglawan 6. Dr. Pisut Niumsup 7. Dr. Dumrong Leenanuruksa
Contents Page Buffalo Reproduction Effect of Different Doses of hCG at AI on Pregnancy Rates of Repeat Breeder Nili-Ravi Buffalo
Muhammad Saleem AKHTAR
362
Pregnancy Rate in Lactating Buffaloes Treated with or without Estradiol after Estrus Synchronization Protocols at Timed AI
Muhammad Saleem AKHTAR
366
Cholesterol Enhances Post-Thaw Semen Quality in Buffaloes (Bubalus bubalis)
Abdul SATTAR
370
Assessment of the Rate of Pregnancy in Buffaloes Crossbred Lactation using Two Protocols CIDR-SYNCH® in 6 and 8 Days
Nestor Simon Montiel URDANETA
375
Increasing Efficiency of Artificial Insemination (AI) in Buffalo Upgrading Program in Nueva Ecija, Philippines
Felomino V. MUMUAD
376
Thyroid Hormone Levels during Oestrous Cycle in Pandharpuri Buffalo
Santosh Hirba DALVI
380
Efficiency of OPU-IVEP-ET of Fresh and Vitrified Embryos in Buffaloes
Wilson SALIBA
385
Pregnancy Monitoring of In Vitro Produced Embryos in Buffaloes
Wilson SALIBA
389
Development of Frozen Buffalo Semen Production in the Phillippines
Felomino MAMUAD
392
Morphology and Biometrics of Spermatozoa of Two Breeds of Water Buffalo (Bubalus bubalis L.)
Sofronio P. KALAW
393
Development of Assessing Motility in Computer Assisted Sperm Analyzer and Phase Contrast Microscopy
Emma VENTURINA
397
A Two-step Culture System Sustains Individual Buffalo Embryo Development
Wenxin ZHANG
401
Comparison of Motility, Morphology, Acrosome Integrity, Membrane Integrity and Fertilizing Ability of Frozen-Thawed Buffalo Sperm Separated by a Percoll® Gradient or PureSperm®
Nutthee AM-IN
405
Effect of Percoll® Density, Duration and Force of Centrifugation on Sperm Motility, Morphology, Acrosome Integrity, Membrane Integrity and Sperm Recovery Rate of Frozen-Thawed Buffalo Semen
Vibuntita CHANKITISAKUL
409
Determination of Pregnancy Associated Glycoproteins Levels in Water Buffalo Cows after Calving
Rafael PAIVA
413
Controlled Breeding and Reproductive Management in Buffaloes using EAZI Breed CIDR
Shivayogayya HIREMATH
418
A Comparison of In Vitro Development of Buffalo (Bubalus bubalis) Embryos Produced either by Somatic Cell Nuclear Transfer or In Vitro Fertilization using Oocytes Obtained by Different Methods
Jianghua SHANG
423
Optimization of Culture and Cryopreservation of Hand-Made Clone (HMC) Buffalo (Bubalus bubalis) Embryos
Jianghua SHANG
427
The Impact of Linolenic Acid on In Vitro Development of Buffalo Embryos
Jianghua SHANG
432
A Preliminary Study on Stable Transfection of EGFP in Buffalo Cumulus Cells
Jianghua SHANG
436
Scrotal Circumference Growth Curves of Buffalo Bulls of Different Breeds Raised in Brazil
Marc HENRY
439
Validation of an ELISA for Detection of Pregnancy Associated Glycoprotein in Water Buffalo Cows
Rafael PAIVA
443
Profiles of Pregnancy-Associated Glycoprotein (PAG) Concentrations during Gestation in Swamp Buffalo
Van Hanh NGUYEN
448
Effect of Povidone-Iodine Uterine Flushing on Insemination Success Rate for Grazing Dairy Buffaloes
Caro B. SALCES
452
Effects of Curcumin on Buffalo Embryonic Development In Vitro
Jiang-Hua SHANG
456
Supplementation of Epidermal Growth Factor into In Vitro Maturation Medium Improves Fertilization Efficiency of Swamp Buffalo Oocytes
Nguyen VIET LINH
460
Effect of Improving Reproductive Management on Dairy Farm Economics
Nasim AHMAD
464
Localization of GnRH Receptors in Buffalo Cow Pituitary Gland in Follicular and Luteal Phases
Thuchadaporn CHAIKHUN
468
Progestin-Based Protocol for Synchronization of Follicular Wave Emergence in Buffaloes during Summer and Winter
Lindsay Unno GIMENES
473
Influence of Parity and Season of the Year on Oocyte Quality and Number in Buffaloes
Lindsay Unno GIMENES
476
Comparison of Two Synchronization Protocols for Timed Artificial Insemination in Acyclic Italian Mediterranean Buffalo Cows out of the Breeding Season
Domenico VECCHIO
479
Morphometric and Functional Characteristics of Buffaloes and Cows Corpus Luteum at Different Stages of the Estrous Cycle
Eunice OBA
480
Response to the First GNRH and Pregnancy Outcome in Buffaloes Underwent Ovsynch and Fixed Timed Artificial Insemination
Gianluca NEGLIA
483
Comparison of Botu-Bov and Tris as Freezing Extenders of Buffalo Sperm Recovered from Epididymal Cauda
Eunice OBA
484
Adding Motility Stimulants to Improve Freezing of Buffalo Sperm Recovered from Epididymal Cauda
Eunice OBA
487
Effect of Testicular Thermoregulation on the Quality of Buffalo Sperm
Carlos Ramires NETO
489
Attenuation of LPS Induced Proinplammatory Gene Expression by Conjugated Linoleic Acid (CLA) in Buffalo Granulosa Cells
Vengala Rao YENUGANTI
493
Frozen-Thawed Epididymal Sperm Quality and the Success Rate of Artificial Insemination in Spotted Buffaloes (Bubalus bubalis carabanensis)
Yusnizar YULNAWATI
494
Supplementation of Buffalo Follicular Fluid: Beware of Other Sources of Steroid Hormones during Culture
Kitiya SRISAKWATTANA
498
Use of Commercially Available Bovine Semen Sexing Agent in Buffalo: Preliminary Report of the Effect on the Conception Rate
Vittoria Lucia BARILE
502
Leptin and Pregnancy: Preliminary Results in Buffalo Cows (Bubalus bubalis)
Vittoria Lucia BARILE
505
Preliminary Evidence on Effect Induced by HeifersPlusTM after In Vitro Exposure on Functional Parameters of Buffalo's Spermatozoa
Vittoria Lucia BARILE
509
Pre-ovulatory Follicle Size, LH Peak Values and Pregnancy Induced by Three Synchronization Treatments in Buffalo Cows, during Non Breeding Season
Giuseppina MariaTERZANO
513
Progesterone Treatment during the Periovulatory Period Decreases Embryo Production in Superovulated Buffaloes
Júlia Gleyci SOARES
517
PGF2α Treatment during the Periovulatory Period Increases the Number of Embryos Recovered from Superovulated Buffaloes
Júlia Gleyci SOARES
522
Use of Different Progestagens for Ovulation Synchronization and TAI in Buffaloes during the Non Breeding Season
Nelcio Antonio Tonizza de CARVALHO
527
Use of Intravaginal Progesterone Devices during Eight or Nine Days in the Ovulation Synchronization Protocol for TAI in Buffaloes during the Non Breeding Season
Nelcio Antonio Tonizza de CARVALHO
532
Plasma Progesterone Radioimmunoassay as a Tool to Confirm Ovarian Cyclicity of Recipients Following Nonsurgical Transfer of Cloned Swamp Buffalo (Bubalus bubalis Lin.) Embryo
Wanvipa SUTHIKRAI
537
Seasonal Effect on Oocytes Recovery Rate and Maturation Rate of Swamp Buffalo Ovaries Collected from Slaughterhouse in Thailand
Kriengsak TASRIPOO
541
In Vitro Embryo Production and Transfer of Bubaline Embryos Using Oocytes Derived from Transvaginal Ultrasound-Guide Follicular Aspiration (TUFA)
Flocerfida AQUINO
538
L-Carnitine Influences the Developmental Competence of Buffalo Oocytes from Aged Donors
Marlon B. OCAMPO
542
ART’s for In Vitro Production of Buffalo Embryos (Philippine Experience)
Marlon B. OCAMPO
543
Developmental Competence of Water Buffalo Oocytes In Vitro in Single Base Medium-Synthetic Oviductal Fluid
Marlon B. OCAMPO
544
Blastocyst Formation after Intracytoplasmic Sperm Injection in Bovine and Buffalo Oocytes Derived from Slaughterhouse
Prudencio B. PEDRO
545
Somatic Cell Nuclear Transfer as a Tool for the Multiplication of Genetically Superior Water Buffaloes: The Philippine Initiatives
Edwin C. ATABAY
549
Improving Ovulation and Conception Rates in Oestrus Synchronized and Artificially Inseminated Water Buffalo Cows by Immunization against Inhibin
Bahareldin-Ali ABDALLA
553
Uterine Microbial Flora of Nili-Ravi Buffalo during Estrus and Its Relationship with Pregnancy Rate in Pakistan
Masood RABBANI
557
Reproductive Performance of Murrah Buffaloes under Intensive Farming System in Thailand
Jamlong MITCHAOTHAI
561
Fetal Loss in Dairy Buffaloes in Eastern of Thailand
Jatuporn KAJAYSRI
565
Study of Frequency Distribution of Calving of River Buffaloes (Bubalus bubalis) in Twelve Farms in Different Ecological Areas in Venezuela
Nestor Simon Montiel URDANETA
568
Generation of Genetically Modified Buffalo Embryos using Multiple-Locus Gene Targeting
Chunying PANG
569
Effect of Frozen Semen from Italian Mediterranean Buffalo on Some Reproductive Parameters and Conception Rate Performed to Different Species Water Buffalo in Southern China
Guangsheng QIN
570
Early Embryo Development in Buffalo
Adriana Caroprezo MORINI
575
Glucose, Cholesterol, Total Protein and Growth Factor Insulin-like Type I in Follicular Fluid of River Buffaloes (Bubalus bubalis)
Carlos GALLEGO
579
Uterine Involution, Fluid Accumulation and Ovarian Activity after Injection of Prostaglandin at Different Periods of Buffalo Puerperium
Eunice OBA
583
Values of 13,14-dihydro-15-keto-PGF2α, Progesterone and Oestradiol after Injection of Prostaglandin at Different Periods of Buffalo Puerperium
Eunice OBA
586
Molecular Evaluation of Developmental Competence of Oocytes Collected In Vivo from Buffalo and Bovine Heifers during Winter and Summer
Laís Mendes VIEIRA
596
The Carabao Development Program as the Cornerstone of the Livestock Biotechnology Program in the Philippines
Eufrocina P. ATABAY
601
Factors Affecting the Performance of an Artificial Insemination Program in North Coast Colombia
Jesus BERDUGO
604
First Report of an Artificial Insemination Program with Sexed Semen in North Coast Colombia
Juan ANGEL
607
Assessment Buffalo Spermatozoa Parameters in using Flow Cytometry
Jesus BERDUGO
609
Effects of L-carnitine Supplemented in Maturation Medium on the Maturation Rate of Swamp Buffalo Oocytes
Teewara PHONGNIMITR
613
Influence of Growth Factors on Survival and Development of Swamp Buffalo Early Antral Follicle Cultured In Vitro
Kwanrudee KAEWMUNGKUN
617
Genetic Parameters for Reproductive Traits of Crossbred Buffaloes from Brazil, Estimated by Bayesian Inference
Carlos Henrique Mendes MALHADO
623
Genetic Parameters for Growth Traits of Mediterranean Buffaloes from Brazil, Estimated by Bayesian Inference
Vanius FALLEIRO
627
Growth Traits of Anatolian and Anatolian x Italian Crossbred Buffalo Calves under the Village Conditions
Özel ŞEKERDEN
632
Population Parameters Based on Known Pedigree Records from Brazilian Jaffarabadi Buffaloes
Carlos Henrique Mendes MALHADO
637
Inbreeding, Average Relatedness Coefficient and Effective Population Size in Jaffarabadi Buffaloes Raised in Brazil
Carlos Henrique Mendes MALHADO
Genetic Parameters for Milk Yield and Lactation Length of Crossbred Buffaloes from Brazil by Bayesian Inference
Carlos Henrique Mendes MALHADO
646
Genetic Correlation for Pre-Weaning and Post-Weaning Traits of Mediterranean Buffaloes from Brazil, Estimated by Bayesian Inference
Vanius FALLEIRO
650
Productive and Reproductive Traits in Murrah Breed from Brazil
Alcides Amorim RAMOS
654
Genomic Exploration of Pakistani Buffalo: Our Black Gold
Masroor Ellahi BABAR
658
Application of Reactions Norms in Study of Genotype Environmental Interaction for Milk Yield of Buffaloes
Humberto TONHATI
662
High Conservation of SRY Gene in Buffalo Compared to Other Bovids
Tanveer HUSSAIN
666
Abstracts Buffalo Genetics and Breeding
641
The Swamp Buffalo: Domestication, Dispersal, and Genetic Differentiation
Yi ZHANG
671
Single Nucleotide Variations in the Buffalo Kappa-Casein Gene (CSN3)
Nedenia Bonvino STAFUZZA
675
Allelic Variant Analysis of the DRB3 Gene by PCR-RFLP in Mediterranean Buffaloes Breed
Nedenia Bonvino STAFUZZA
680
Cattle-Derived Microsatellite Markers to Study the Water Buffalo Genome
Nedenia Bonvino STAFUZZA
681
Genetic Estimates for Productive Parameters in Buffaloes in Different Ecological Areas in Venezuela
Nestor Simon Montiel URDANETA
686
Index of Heritage's Weight at Birth and Effects of Some Environmental Factors on Buffalo Calf using the Animal Model
Nestor Simon Montiel URDANETA
687
Genetic Parameters for Productive and Reproductive Traits for Milk Buffalo in Brazil
Camila da Costa BARROS
688
Genetic Variants of POU1F1 Gene in Azakheli Buffalo Breed of Pakistan
Asif NADEEMA
692
Single Nucleotide Polymorphisms in Bovine CYP11B1 Gene in Nili Ravi Buffalo Breed of Pakistan
Maryam JAVED
697
Identification of Single Nucleotide Polymorphisms in OLR1 Gene in Nili Ravi Buffalo
Maryam JAVED
701
POU1 Transcription Factor 1 DNA Polymorphism in Nili Ravi Buffalo
Rashid MAJEED
706
Bovine Stearoyl-CoA Desaturase Gene Polymorphism in Nili Ravi Buffalo
Maryam JAVED
710
Genetic Parameter Estimates for Milk Yield and Lactation Length in Buffalo
Yenny GARCÍA
714
Genetic Parameters and Trends for Weaning Weight and Calving Interval of Department of Livestock Development Swamp Buffalo
Nikorn SANGHUAYPHRAI
717
Genetic Polymorphism in Caspase Activating Recruitment Domain 15 and Its Association with Incidence of Mastitis in Murrah Buffalo
Archana VERMA
723
Genetic Variability in Production and Immune Function Genes Associated with Production Traits and Incidence of Mastitis in Indian Murrah Buffalo
Archana VERMA
729
Genotyping and Molecular Characterization of NRAMP1/-2 Genes as Location of Markers for Resistance and/or Susceptibility to Mycobacterium bovis in Swamp and Riverine Type Water Buffaloes
Lawrence P. BELOTINDOS
730
Buffalo Genetic Improvement Programme in Nepal-Current Status and Future Prospects
Bhola Shankar SHRESTHA
734
Optimization of the Transfection Efficiency of Buffalo Fetal Fibroblasts Cells using Liposome
Tingxian DENG
744
The Construct of Fat-1 Gene Targeted Buffalo Kidney Fibroblast Cell
Yingyin ZHANG
745
Multiple-trait Genomic Evaluation for Milk Yield and Milk Quality Traits using Genomic and Phenotypic Data in Buffalo in Brazila
Humberto TONHATI
746
Preliminary Results on the Growth Performance of F1 Mediterranean Buffalo Offspring Crossed in China
Guangsheng QIN
750
Karyotype Analysis of Mediterranean Buffalo and Its Hybrids
Fenxiang HUANG
755
Estimation of Genetic Parameters for Growth Traits of Three Genotypes of Water Buffalo Bulls Raised on a Ranching Operation
Agapita J. SALCES
760
Some Environmental Factors Affecting Performance Traits in Registered Nili Ravi Buffalo Population at Field Area of Pakistan
Khalid JAVED
764
Genetic Parameters for Milk Yield and Milk Component Traits Estimated from Test Day and 305D Lactation Records of Philippine Dairy Buffaloes
Ester B. FLORES
768
Improvement, Utilization and Conservation of Buffaloes under New Legislation of Pakistan
Muhammad Nawaz SAEED
773
Genetic Parameters for Test-Day Fat Yield Estimated by Random Regression Models in Dairy Buffaloes using Bayesian Inference
Humberto TONHATI
774
Development of Multiplex PCR for Sexing Buffalo Embryos
Wisut NUALCHUEN
775
Studies on Linear Type Traits and Morphometric Measurements in Nili Ravi Buffaloes of Pakistan
Khalid JAVED
780
Identification of Amino Acid Substitutions and Structural Model Prediction of POU1F1 Gene in Azakheli Buffalo Breed of Pakistan
Asif NADEEM
784
Polymorphisms in Osteopontin Gene in Amazon Buffaloes
Sebastião Tavares ROLIM FILHO
788
Relationship between Body Measurements and Milk Production in Nili-Ravi Buffaloes Maintained at Commercial Farms in Peri-urban Vicinity of Lahore
Nisar AHMAD
792
Buffalo Milk Transcriptomics
Christophe LEFEVRE
796
Compare of Phenotypic Variation with the Polymorphism of One Coagulation Related Gene Locus in Buffalo KappaCasein
Qingkun ZENG
805
Comparative Proteomic Analysis of the Changes of Milk Protein Associated with Different Breeds of Buffalo
Qingkun ZENG
811
Productive and Reproductive Parameters in Buffaloes and Cows in a Farm Located in Tropical Dry Forest in Venezuela
N. MONTIEL-URDANETA
812
Productive and Reproductive Parameters in Buffaloes and Cows in a Farm Located in Tropical Humid Forest in Venezuela
N. MONTIEL-URDANETA
813
Microsatellite Analysis of Thai Swamp Buffalo Cloned Calf Derived from Ear Fibroblasts
Supitchaya TREEBONMUANG
814
The Comparison of Some Reproductive Traits of Anatolian and F1 Crossbred (Anatolian X Italian) Buffalo under Village Conditions in Turkey
Özel ŞEKERDEN
819
Minerals Status of Soil, Fodder and in Lactating Nili-Ravi Buffaloes in Irrigated Agro-ecological Zone of Punjab, Pakistan
Muhammad Saleem AKHTAR
824
Digestibility and Performance of Buffalo Fed Total Mixed Ration with Different Levels of Citric Waste
Suthipong URIYAPONGSON
829
Milk Yield Response of Bypass Protein Feeding (Soybean Meals) in Dairy Animals
Netra Prasad OSTI
834
Body Composition and Net Energy Requirements for Maintenance of Non-Castrated Water Buffaloes (Bubalus bubalis)
André Mendes JORGE
840
Allometry of Organs and the Gastrointestinal Tract from Water Buffaloes (Bubalus bubalis) Finished in Feedlot
André Mendes JORGE
844
Distribution of Beta-Catenin in Colon Precarcinomatous Lesions of Rats Fed with Functional Buffalo Milk
Marcial SÁNCHEZ NEGRETTE
848
Anticancer Effects of Bubaline Functional Milk with Higher Concentration of Conjugated Linoleic Acid and Omega-3 Fatty Acids
Gabriela Verónica RAMIREZ
853
Virtues of the Milk from Water Buffalo
Muhammad YOUNAS
857
Improving Oat Grass Silage Quality through using Exogenous Enzyme in Cannulated Buffalo Bulls
Mahr-un-NISA
866
Effects of Roughage Types on Feed Intake, and Nutrient Digestibilities in Swamp Buffaloes (Bubalus bubalis)
Walailuck KAEWWONGSA
871
Comparative Performance of Calves Fed Milk and/or Milk Replacer Supplemented with Calf Starter up to Weaning Age in Nili-Ravi Buffaloes
Muhammad ABDULLAH
874
Changes in Molecular Diversity of Rumen Methanogens in Buffalo and Cattle in Response to Dietary Tannin
Gondelina A. RADOVAN
878
Abstracts Buffalo Nutrition and Feeding
Comparative Utilization of Different Types of Roughage in Thai Swamp Buffalo and Thai Brahman Cattle Based on In Vivo Nutrient Utilization, Nitrogen Balance and Purine Derivatives Excretion in the Urine
Thongsuk JETANA
883
454 GS FLX Pyrosequencing Reveals Rumen Bacterial Diversity of Chinese Water Buffalo
Chengjian YANG
888
Rumen Bacterial Diversity of Murrah and Nili-Rivi Buffalo (Bubalus bubalis) Assessed by 454 GS FLX Pyrosequencing
Chengjian YANG
889
Novel Methods to Improve the Nutritive Value of Low Quality Roughages for Nili Ravi Buffalo Calves
Faisal SHAHZAD
890
Effect of Cysteamine Hydrochloride on In Vitro Methane Emission in Water Buffalo
Caixia ZOU
894
Evaluation of Nutritive Value of Agricultural By-Products and Industrial By-Products for Buffalo by Cornell Net Carbohydrate and Protein System
Junhua ZHOU
895
Effects of Beer Lees and Cassava Residues Respectively Substituting for Soybean Meal and Grassiness on Milk Performance in Lactating Water Buffalo
Caixia ZOU
900
Graded Replacement of Corn Grain by Wheat Grain in Sahiwal Calves: Influence on Nutrients Intake, Digestibility, Blood Metabolites and Growth Performance
Muhammad Aasif SHAHZAD
904
Response and Ruminal Characteristics of Buffalo Bulls Fed Urea-Molasses Treated Wheat Straw Inoculated with Rumen Digest
Muhammad SARWAR
908
Response of Sahiwal Heifers Receiving Maize Fodder with Supplementation of Urea Molasses Block
Nasir Ali TAUQIR
913
Performance of Nili Ravi Buffalo Calves Fed Urea-Corn Steep Liquor Treated Corn Cobs
Muhammad Aasif SHAHZAD
918
Estimation of In vitro Methane Production in Buffalo and Cow
Federico INFASCELLI
924
Chemical Composition, Rumen Fermentation Kinetics, Digestibility and Energy Value of Cassava Leaves Hay at Different Storage Times
Patricia. M. Guimaraes BEELEN
928
Effects of substituting Beer Lees and Cassava Residues Respectively for Buffalo Dietary Soybean Meal and Grassiness on Rumen Fermentation In Vitro
Zhongsheng XIA
929
Nutritive Value and In Situ Digestion Kinetics of Some Leguminous and Non-leguminous Fodder Baled Silages in Buffalo Bulls
Nawaz SAEED
938
Potential Benefits from the Utilization of Some Natural Feed Resources in Thai Swamp Buffaloes
Thongsuk JETANA
942
Effects of Lasia spinosa Thw. and Season on Plasma Leptin and Glucose of Weaned Female Murrah X Swamp Buffalo Calves
Ratree JINTANA
947
Rumen Bacterial Diversity of Water Buffalo (Bubalus bubalis) as Influenced by Concentrate Levels
Chengjian YANG
951
Effect of Rumen-Protected Methionine and Reduced Crude Protein in Lactating Mediterranean Buffaloes Diet
Antonella CHIARIOTTI
952
Effect of Varying Levels of NDF on Voluntary Intake, Nutrients Digestibility for Nili Ravi Buffalo Heifers
Saeed AHMED
957
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
Daniel Lopez AQUINO
961
Effect of Protein on Microbial Protein Synthesis and Productive Performances of Thai Swamp Buffalo (Bubalus bubalis)
Pramote PAENGKOUM
966
Effect of Protein Level and Urea Source in Concentrate on Feed Intake and Rumen Ecology in Swamp Buffalo Fed Rice Straw
Sungchhang KANG
970
Effect of Dried Leucaena Leaf Supplementation on Rumen Ecology, Nutrient Digestibility and Urinary Excretion of 2,3Dihydroxy Pyridone (2,3-DHP) and 3,4-Dihydroxy Pyridone (3,4-DHP) in Swamp Buffaloes
Kampanat PHESATCHA
975
Effects of Eucalyptus Crude Oils Supplementation on Nutrients Digestibility of Swamp Buffaloes
Nguyen The THAO
980
Influence of Urea-Calcium Mixture in High-Quality Feed Block on Ruminal Fermentation in Swamp Buffalo
Anusorn CHERDTHONG
984
Nutritional Status of Some Trace Minerals of Water Buffaloes in Egypt
Maha Mohamed HADY
988
Effect of Roughage Sources and Fibrolytic Enzyme Supplementation on Nutrient Digestion and Rumen Fermentation in Buffaloes
Chalermpon YUANGKLANG
993
Antigen Based Detection of Cystic Echinococcosis in Buffaloes using ELISA and Dot – EIA
Arumugam SANGARAN
999
Comparative Efficacy of Enrofloxacin and Oxytetracycline as Systemic Dry Period Therapy for the Control of Bubaline Mastitis
Muhammad KASHIFA
1002
Incidence and Organ Wise Involvement of Hydatidosis in Buffaloes
Arumugam SANGARAN
1009
Buffalo Health
Emphysemated Necrotic Skin Disease (Patakha): a Newly Emerging Disease in Buffaloes
Abdul SHAKOOR
1011
Comparative Efficacy of Three Indigenous Plants (Fumaria parviflora, Artemesia maritima & Swertia chirata) Alone or in Combination for the Treatment of Toxaemia
Sayyed AUN MUHAMMAD
1016
In Vivo Comparison of Specific Activity of Egg Yolk Immunoglobulins (IgY) and Antibiotic Against Staphylococcus aureus Causing Mastitis in Buffaloes (Bubalus bubalis)
Tanveer AHMAD
1017
Antibiogram Analysis of Staphylococcus aureus isolated from Mastitic Milk Samples of Buffaloes in District Bhimber Azad Kashmir
Abid HUSSAIN
1021
Molecular Identification of Brucella abortus Bv5 and Strain 19 in Water Buffaloes (Bubalus bubalis) in Northeast Argentina
Diana MARTINEZ
1029
Efficacy Analysis of Parasitic Integrated Control in Buffaloes
Eduardo BASTIANETTO
1033
Detection of Toxin Genes by PCR in Clostridium perfringens Isolates Collected from Water Buffaloes (Bubalus bubalis) Affected by Lethal Enterotoxemia
Esterina DE CARLO
1035
BUN and Total Protein Levels of Buffalo Population in Udonthani Province of Thailand
Chanachai BOONPERM
1038
Prevalence and Molecular Diagnosis of Staphylococcus aureus Subclinical Mastitis in Lactating Nili-Ravi Buffaloes (Bubalus Bubalis) at Livestock Experiment Station, Bahadurnagar, Okara, Pakistan
Waseem SHAHZAD
1041
Prevalence, Molecular Diagnosis and Treatment of Field Isolates of Toxogenic Pasteurella multocida in a Hemorrhagic Septicemia Outbreak in Nili-Ravi Buffalo Calves at Livestock Experiment Station, Bahadurnagar, Okara, Pakistan
Waseem SHAHZAD
1046
Management of Khari Disease Syndrome with Pentasulfates Supplementation in Lactating Buffaloes
Doj RAJ KHANAL
1051
Isolation of Mycoplasma capricolum subspecies capricolum from Dairy Buffalo (Bubalus bubalis)
Esterina DE CARLO
1056
Bubaline Herpesvirus 1 Associated with Abortion in a Mediterranean Water Buffalo
Esterina DE CARLO
1059
Doramectin Resistance in Helminths from Buffaloes
Euardo BASTIANETTO
1063
Breeding Techniques, Welfare and Mammary Gland Pathologies in Buffalo
Esterina DE CARLO
1066
Comparison between Two Gamma-Ifn Assays and Intradermal Tuberculin Test for the Diagnosis of Tuberculosis in Water Buffalo (Bubalus bubalis)
Esterina DE CARLO
1071
Carrier Status of Foot and Mouth Disease in Ruminants through Reverse Transcription Polymerase Chain Reaction
Muhammad Hassan MUSHTAQ
1075
The Metaphylactic Efficacy of Toltrazuril (Baycox® Bovis Bayer) and Diclazuril (Vecoxan® - Esteve Veterinaria) in Natural Infections of Eimeria spp. in Buffalo Calves: a Multicenter Trial in Southern Italy
Giovanna CAPPELLI
1076
Gastrointestinal Parasitic Diseases of Buffaloes and Implications of Climate Change for these Diseases in Nepal
Bhoj Raj JOSHIP
1082
Detection of Bovine Viral Diarrhea Virus Prevalent in Dairy Herds of Punjab, Pakistan
Masood RABBANI
1088
Clinical Evaluation of Hypertonic Saline Solution for Treatment of Lactic Acidosis in Water Buffaloes
Frederico Augusto Mazzocca Lopes RODRIGUES
1091
Influence of Storage Temperature on Blood Gas Analyses on Buffalo Venous Blood
Antonio Humberto Hamad MINERVINO
1092
Experimental Induction of Lactic Acidosis in Buffaloes with Sucrose
Raimundo Alves BARRÊTO-JÚNIOR
1093
Mineral Status of Buffaloes Raised in the Wetlands of the Lower Amazon Basin Ecosystem
Enrico Lippi ORTOLANI
1094
Serological Profile of Buffalo (Bubalus bubalis) Female Calves Vaccinated with Standard Brucella abortus Strain 19 Vaccine using Rose Bengal, 2-Mercaptoethanol and Complement Fixation Tests
Geraldo de NARDI JÚNIOR
1095
Interference of Vaccinal Antibodies on Serological Diagnostic of Leptospirosis in Vaccinated Buffalo using Two Types of Commercial Vaccines
André Mendes JORGE
1099
Vaccine Trial of Recombinant Schistosoma japonicum Paramyosin in Water Buffaloes
Mario Antonio L. JIZ II
1103
Seroepidemiological Investigation, Risk Factors Analysis of Brucellosis in Ruminants and Their Owners in District Buner of Pakistan
Mansur-ud-Din AHMAD
1104
Disease Susceptibility of Buffalo in Pakistan
Sidra MANZOOR
1110
Epidemiological Studies on Mastitis in Pakistani Buffalo
Sidra MANZOOR
1114
Excretion of Aflatoxin M1 in Milk of Mediterranean Italian Buffalo Cow Fed Diet Naturally Contaminated of Aflatoxin B1
Carlo BOSELLI
1119
Preliminary Results on the Utilization of Phytotherapy in Organic Italian Mediterranean Buffaloes
Domenico VECCHIO
1123
Buffalo Physiology Bone Marrow´s Harvest in the Coxal Tuberosity for Isolation and Culture of Mesenchymal Stem Cells of Buffaloes (Bubalus Bubalis)
Eunice OBA
1125
Isolation, Culture and Differentiation of Buffaloes Bone Marrow Mesenchymal Stem Cells Obtained from the Coxal Tuberosity
Eunice OBA
1128
Haematochemical and Hormonal Parameters Related to Buffalo Calves Welfare
Cristina RONCORONI
1131
A Preliminary Study about Lymphocyte Subset of Water Buffalo Calves
Cristina RONCORONI
1136
Impact of Thermal Stress on Rectal, Skin Surface Temperatures, Respiration Rate, Heat Load Index and Heat Storage in Lactating Murrah Buffaloes (Bubalus bubalis)
Vijay KUMAR
1141
Scrotal Thermography and Doppler Ultrasonography of the Testicular Artery of Buffaloes Subjected to Environmental Heat Stress
Carlos Ramires NETO
1145
Blood Biochemical Profiles of Mehsana Riverine and Thai Swamp Buffaloes under Tropical Conditions in the Northeast Thailand
Suvit BOONPRONG
1146
Changes in Insulin-Like Growth Factor-Binding Proteins in Swamp Buffalo Uterus during Estrous Cycle
Sirima THONGRUAY
1151
Use of Oxytocin and Milking Management of Buffaloes in (Urban) Peri-Urban Area of Faisalabad
Muhammad TARIQ
1155
Environmental Factors Affecting Live Weight and Morphological Traits in Nili Ravi Buffaloes of Pakistan
Khalid JAVED
1161
BUN and Total Protein levels of Buffalo Population in Udonthani Province of Thailand
Chanachai BOONPERM
1165
Preliminary Study on Energy Metabolism and Energy Requirement of Early Lactation Water Buffalo
Caixia ZOU
1168
Inter-relationship of Milk Constituents with Body and Udder Measurements in Nili-Ravi Buffaloes Raised at Commercial Farms of Pakistan
Khalid JAVED
1170
An Integrated Study on Milk and Beef Production Conducted at Macún Buffalo Enterprise in Cuba, Some Results and Recommendations
Luis Mateo FRAGA BENITEZ
1174
Buffalo Production and Management
Productivity of Mehsana Riverine Buffalo under Tropical Conditions of Thailand
Prapawan SAWASDEE
1175
Status and Perspectives of Buffalo in Bangladesh
Quazi M EMDADUL HUQUE
1179
Buffalo Farming in Bhutan: Challenges & Opportunities
Nar Bahadur TAMANG
1184
Transition of Milk Production and Reproduction of Dairy Buffaloes in Nepal
Yoshiaki HAYASHI
1188
Community Management for Buffalo Eco-Tourism in Udon Thani
Krisdakorn WONGWAI
1193
Economical and Social Importance of Buffalo in a Small Milk Production System
Eduardo BASTIANETTO
1197
Coordination of the Chain of Buffalo Milk in São Paulo State (Brazil)
Fabrício Pini ROSALES
1200
Swamp Buffalo Production System and Needs for Extension on Local Scale Farmers in the Lower Northeast of Thailand
Sornnarong SUPHACHAVALIT
1204
The Problems and Obstacles on Raising Buffaloes of Local Farmers in Central Thailand: A Case Study of Saraburi Province
Sirisuk SAPAPANAN
1208
Assessment of Village-Based Artificial Insemination Technician (VBAIT) Scheme as a Strategy Towards Privatization of Artificial Insemination (AI) Services in Nueva Ecija
Sonia D. POL
1212
Condition and Specifics of Buffalo Breeding in Republic of Macedonia
Bone PALASHEVSKI
1218
Assessing the Performance of Village-Based Artificial Insemination Technicians for Water Buffaloes in Nueva Ecija Province, Philippines
Eric PALACPAC
1222
Sustainability of Philippine Carabao Center and Primary Cooperative Partnership in Carabao-Based Enterprise
Wilma T. Del ROSARIO
1226
A Model of Sustainable Herd Buffalo Farming in Songkhram Wet Land, Nakhon Phanom
Tanapat SURANARAKUL
1230
Contextualizing the Gatasng Kalabaw Festival in Support to the Carabao-based Enterprise in Nueva Ecija, Philippines
Marlowe U. AQUINO
1233
Economics of Raising Calves on Milk and/or Milk Replacer in Nili-Ravi Buffaloes
Muhammad SAADULLAH
1237
Biohydrogen Production from Buffalo Manure Codigested with Agroindustrial By-products in an Anaerobic Reactor
Antonella CHIARIOTTI
1241
Buffalo Socio-economic and Sustainable Production
The Herd Size and Production Performances of Buffalo in Romania
Livia VIDU
1245
Fatty Acids in the Muscle and Fat Layer of Buffaloes Supplemented with Fish Oil
Exequiel Maria PATIÑO
1250
Using Slaughter Weight to Predict Weight and Yield of Primal Cuts of Carcass from Buffaloes
André Mendes JORGE
1254
Carcass Composition and Meat Quality of Buffalo by Raised Alongside Mekong River: Nakhon Phanom Province
Tanom TATHONG
1258
Effect of Gender on Carcass Composition and Meat Quality of Buffalo in Wet-Land in Nakhorn Phanom
Tanom TATHONG
1262
Tenderization Effect on the Physical Characteristics of Commercial Cuts of Second and Third Buffalo´s Meat (Bubalus bubalis) during Ripening Process
Laura Marcela MENDOZA ARAGÓN
1266
Conjugated Linoleic Acid and Fatty Acids Profile in Buffalo Meat
Federico INFASCELLI
1270
Determination of Muscularity and Correlation with Body Weight in Buffalo in the Northeast of Argentina
Gladis REBAK
1274
Relation of the Approximate Age and Ultrasound Data in buffalo in the Northeast of Argentina
Gladis REBAK
1277
Research regarding the dynamics of body development in young buffaloes, depending on various factors
Livia VIDU
1280
Estimation of Fine Cuts of Meat Production Performance in Mediterranean Italian Buffalo Young Bulls
Roberta VITTORIA
1284
Body Condition Score (BCS) System in Murrah Buffaloes
Kappa Sarjan RAO
1290
Flavored Probiotic (Acidophilus) Buffalo Milk: Development and Quality Assessment
Muhammad JUNAID
1300
Effective Environmental Factors on Milk Composition, Rennet Coagulation Time and Urea Content of in Anatolian Buffaloes Milk of Ilikpinar Village Hatay Province
Özel ŞEKERDEN
1305
A Study on the Composition and Microbiology of Raw Milk from Three Breeds of Buffalo in Thailand
Jitkamol THANASAK
1311
Quality Evaluation of Olive Oil Coated Labneh Cheese Mixed with Culinary Herbs
Sarfraz AHMAD
1316
Seasonal Variations in Chemical Composition of Buffalo Milk
Sarfraz AHMAD
1324
Federico INFASCELLI
1330
Buffalo Meat and Meat Products
Buffalo Milk and Milk Products
Milk CLA Content and ∆9 Desaturase Activity in Buffalo Cows along the Lactation
Milk Yield and Milking Characteristics in Murrah Buffaloes Submitted to Machine Milked with or without Calf
Alberto de GUSMÃO COUTO
1334
Relationship of Udder and Teat Morphology with Milk Production in Nili-Ravi Buffaloes of Pakistan
Muhammad ABDULLAH
1335
Differences in Lactation Curves and Peak Production in Mediterranean Italian Buffaloes Bred According to Three Levels of Production
Angelo COLETTA
1339
Microstructure, Rheological and Textural Characteristics of Low Fat Buffalo Milk Cheddar Cheese
Mian Anjum MURTAZA
1346
Buffalo Bulletin 2013 Vol.32 (Special Issue 2): 564-567
Uterine Microbial Flora of Nili-Ravi Buffalo during Estrus and its Relationship with Pregnancy Rate in Pakistan Sohail RAZA, a Masood RABBANI, a* Nasim AHMAD, b Ali Ahmad SHEIKH, a Khushi MUHAMMAD, c Fareeha AKHTARa and Habib Ur REHMANa a
University Diagnostic Lab, University of Veterinary and Animal Sciences Lahore 54000 Pakistan. Department of Theriogenology, University of Veterinary and Animal Sciences Lahore 54000 Pakistan. C Department of Microbiology. University of Veterinary and Animal Sciences Lahore 54000 Pakistan. *Corresponding email:
[email protected] b
ABSTRACT Microbial flora of body plays a pivotal role in protecting body against infection and enhances its ability to compete pathogens. Some of the microorganisms are beneficial for the uterine body and some are detrimental. Presence of Actinomyces pyogenes and Fusobacterium necrophorum are reported to be important pathogens causing metritis with lower conception rate and increase calving intervals. In order to check the presence of microbes in Nili Ravi buffaloes and its affect on the pregnancy rate, a trial was conducted on 50 buffaloes from an organized dairy farm. Samples were collected using specially prepared devise just before artificial insemination and transferred to the lab for bacterial identification. Two to three months later, pregnancy of the sampled animals was checked. More than 173 strains belonging to six different genera identified as Escherichia coli (E. coli), Lactobacillus spp., Micrococcus spp., Staphylococcus spp., Citrobacter spp. and Proteus spp. were identified. Among these, E. coli was most prevalent (100 %) followed by S. aureus (98 %), Lactobacillus (86 %), Micrococcus (26 %), S. epidermidis (14 %), Proteus (12 %), Citrobacter (10 %). Relationship of bacterial species with pregnancy rate then found that 4 animals aborted in first trimester were having significantly high number (p< 0.05) of Citrobacter species. It is concluded that E.coli, Staph. aureus, S. epidermidis and Lactobacillus species are normal microbial flora of Nili-Ravi buffalo Proteus species and Micrococcus species are mostly present in pregnant animals while presence of Citrobacter species is detrimental for the pregnant animals and may lead to abortion. Keywords: Bio-film, Uterine flora, Nili-Ravi, Pregnancy rate. INTRODUCTION Microbial flora of body plays a significant role in protection against infectious diseases (Reid et al., 1998). Various bacteria can be isolated from uterine body of normal cyclic animals which includes Escherichia coli, Staphylococcus spp., Bacillus spp., Yersinia enterocolitica, Citrobacter diversus, Bacillus spp., Micrococcus spp. (Ahmed et al., 2007; Azawi et al., 2007; El-Jakee et al., 2008 and Gani et al., 2008). Lactobacillus spp. proved beneficial while Actinomyces pyogenes and Fusobacterium necrophorum causes metritis and other uterine diseases (Azawi et al., 2008). These pathogens cause low fertility, hormonal imbalance and stoppage of ovulation (Sheldon et al., 2004). Uterine infections in buffalo are much higher as compare to cows (Jainudeen, 1986). Keeping in view, this study was designed to investigate microbial flora of uterus in buffalo during estrus and to evaluate their impact on pregnancy rate. MATERIALS AND METHODS Sampling A total of 50 uterine fluid/secretion samples of Nili-Ravi buffaloes during estrus period were collected using sterilized artificial insemination (AI) rod fitted with disposable syringe as per protocol Accepted April 10, 2013; Online February 24, 2014. 564
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mentioned earlier (Williams et al., 2007). The samples were transported to University Diagnostic Lab, University of Veterinary & Animal Sciences, Lahore, Pakistan for further processing. A complete clinical history and management of each animal was also recorded. All animals were dewormed and vaccinated as per schedule. Isolation and Identification of the bacteria Primary isolation of the bacteria was carried using blood agar. Bacterial colonies were separated and identified biochemically following instructions to Burgey’s Manual. Bacterial confirmation was also done using API Kits. Pregnancy rate Rectal palpation technique for pregnancy status of these animals was also performed after 2-3 months post insemination. Statistical analysis Data was tabulated using Microsoft Excel (MS Excel 2007, Microsoft Corporation, Redmond, WA). Descriptive Statistical analysis like percentages, confidence interval and graphs were also developed using commercially available statistical package SPSS (Version 18, for Windows; SPSS, Chicago, IL). Statistical association of pregnancy status between the presence and absence of certain isolated micro flora was calculated by using Fisher Exact Test using SPSS. RESULTS AND DISCUSSIONS All uterine samples (100 %) were positive for presence of various bacteria. A total of 173 bacterial isolates belonging to six genera such as E. coli, Lactobacillus spp., Micrococcus spp., Staphylococcus spp., Citrobacter spp. and Proteus spp. were recovered (Table-1). The most prevalent bacterial isolates in both pregnant and non pregnant animals were E. coli, Lactobacillus spp., and S. aureus, while Citrobacter spp. were prevalent in aborted buffaloes. S.epidemidis, Micrococcus spp. and Proteus spp. were prominently observed in the pregnant animals than non pregnant. Similar results were observed by other researchers who have identified E. coli, Staphylococcus spp., Bacillus spp., Yersinia enterocolitica, C. diversus, Bacillus spp., Micrococcus spp. from uterus (Ahmed et al., 2007; Azawi et al., 2007; El-Jakee et al., 2008 and Gani et al., 2008). E. coli (100%) was recovered from pregnant, non pregnant and aborted animals. E. coli is the most commonly isolated bacteria from bovine uterus of normal and puerperal animals (William et el., 2007). E. coli was isolated from normal pre-pubertal buffaloes (Torres et al., 1997 and El-Jakee et al. 2008). Hanafi et al. (2008) studied that prevalence of E. coli in healthy animals was 71. 21% and in endometric animals it was 18.75%. These studies further supported the results of current research that E. coli present in both healthy and diseased animals. Statistical analysis reveals that there is no significant (p=0.9) relationship in pregnancy rate with presence of E. coli in uterus. Prevalence of S. aureus in this study was recorded 98% with no significant relationship with pregnancy rate statistically (p=0.84). Previous findings also indicate that S. aureus, being opportunistic organism, is the most common bacterial isolates of Nili Ravi buffaloes and usually associated with endometritis (Usmani et al., 2000 and William et al., 2007). Prevalence of Lactobacillus spp. and Micrococcus spp. in this study was 86% and 26%, respectively with no statistically significant relationship with pregnancy rate (p=0.289 and p=0.117, respectively). Prevalence of both organisms was more in pregnant animals as compare to non pregnant animals. Previous study also indicates the high prevalence of both of these organisms in normal animals than in animals with ovarian inactivity (Ahmad et al., 2007, Abd El-Moeez et al., 2008 and Hanafi et al., 2008). The isolation of S. epidermidis and Proteus spp. was almost same prevalence from pregnant and non pregnant animals in this study. Ahmad et al., 2007 concluded that S. epidermidis is mostly isolate from pregnant animals. Proteus spp. is most prevalent organism of the normal purpeural animals. Proteus spp. is an opportunistic contaminant of uterus not usually associated with endometritis (Jadon et al., 2005). Prevalence of Citrobacter spp. was 10% in total samples, out of which 80% was present in aborted 565
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animals and 10% in non-pregnant animals. Previous study suggested that Citrobacter spp is co-related with the presence of severe endometrial lesions (Messier et al., 1984). Citrobacter spp. causes the sporadic abortion in bovines which is also evident from current study as there exist a significant relation (p=0.0001) between presence of Citrobacter spp. and pregnancy rate. After insemination, 68% of the animals became pregnant, 24% remained non pregnant and 8% were aborted. Reproductive disease history showed that 30 animals had encountered the reproductive disease. Eight animals had the problem of anoestrus, out of which seven became pregnant and one remain non pregnant. Thirteen animals had the problem of metritis, out of which 3 became non pregnant, one aborted and 9 became pregnant. Six animals had the problem of inactive ovaries, out of which 5 became pregnant and one remained non pregnant. Two animals had the problem of pyometra, out of which one became non pregnant and other aborted. One non-pregnant animal showed the problem of retained placenta. It has been observed in this study that most of the animals which had problem of anoestus became pregnant. Results of this study show that animals which had previous problem of metritis (25 %) were non pregnant while 7.6 % aborted and 68.4 % became pregnant. In the animals having history of inactive ovaries, 16.6 percent became non pregnant while 83.34 % became pregnant and the animals showing the problem of pyometra before, 50 % of them became non pregnant and 50 % aborted. Animal which had the problem of retained placenta remained non pregnant. CONCLUSIONS From the current study, it was concluded that Citrobacter spp.may be linked with problems related to reproductive system of Nili-Ravi buffalo. Furthermore anoestrus, metritis and inactive ovaries are not fatal for the uterine health. Pyometra and retained placenta diseases are most fatal for the reproductive health of the animals especially in Nili-Ravi buffaloes. As this is a preliminary study for the isolation of bacteria from uterus of Nili-Ravi buffalo, there is a need to establish metagenomic study to figure out other microbiotas which are difficult to isolate using conventional isolation techniques and relate them with pregnancy rate of this precious animal. REFERENCES Abd El-Moeez, S.I., W.M. Ahmed, J. A. El-Jakee and F.R. El-Seedy. 2008. Observation on lactobacillus spp. In the genetal tract of buffaloe-cows with emphasis on its in vitro probiotic activity. Global Veterinaria 2(1): 15-21. Ahmed, W.M, J.A. El-Jakee, F.R. El-Seedy, K.I. El-Ekhnawy and S.I. Abd El-Moez. 2007. Vaginal bacterial profile in buffalo-cows in relation to ovarian activity. Global Veterinaria 1(1): 0108. Azawi, O. I., S. N. Omran and J. J. Hadad. 2007. Clinical, Bacteriological and Histopathological Study of Toxic Puerperal Metritis in Iraqi Buffalo. J. Dairy Sci. 90: 4654-4660. Azawi, O.I., M.A. Rahawy and J.J. Hadad. 2008. Bacterial Isolates Associated with Dystocia and Retained Placenta In Iraqi Buffaloes. Reprod. Dom. Anim. 43: 286–292. El-Jakee, J.A., W.M. Ahmed, F.R. El-Seedy. and S.I. Abd El-Moez. 2008. Bacterial profile of the genital tract in female buffaloes during different reproductive stages. Global. Veterin. 2 (1): 714. Gani, M.O., M.M. Amin, M.G.S. Alam, M.E.H. Kayesh, M.R. Karim, M.A. Samad and M.R. Islam.2008. Bacterial flora associated with repeat breeding and uterine infections in dairy cows. Bangl. J. Vet. Med. 6(1): 79-86. Hanafi, E.M., W.M. Ahmed, S.I. Abd-El-Moez, H.H. El-Khadrawy and A.R. Abd-El-Hameed. 2008. Effect of Clinical Endometritis on Ovarian Activity and Oxidative Stress Status in Egyptian Buffalo-Cows. Ameri-Euras. J. Agric & Environ Sci. 4(5): 530-536.
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Jadon, R.S., G.S.Dhaliwal, S.K. Jand. 2005. Prevalence of aerobic and anaerobic uterine bacteria during peripartum period in normal and dystocia-affected buffaloes. Animal Rep. Sci. 88: 215–224. Jainudeen, M. R. 1986. Reproduction in water buffalo. In: Morrow DA (ed), Current Therap in Therio. W.B. Saunders, Philadelphia, PA, pp: 443–449. Messier, S., R. Higgins, Y. Couture and M. Morin.1984. Comparison of Swabbing and Biopsy for Studying the Flora of the Bovine Uterus. Can. Vet J. 25: 283-288. Reid, B.A., A.W. Bruce and V. Sameianov. 1998. The role of Lactbacillus in preventing urogenetal and intestinal infections. Int. Dairy J. 8: 555-562. Sheldon, I.M. and H. Dobson 2004. Postpartum uterine health in cattle. Ani. Rep. Sci. 82-83: 295-306. Torres, E. B., T. Nakao, T. Hiramune, M. Moriyoshi, K. Kawata and K. Nakada. 1997. Stress and uterine bacterial flora in dairy cows clinically normal and abnormal puerperium. J Rep & Dev 43(2): 157-163. Usmani, R.H., N. Ahmad, P. Shafiq and M.A. Mirza. 2000. Effect of subclinical uterine infection on cervical and uterine involution, estrus activity and fertility in postpartum buffaloes. Theriogenology 55: 563-571. Williams, E.J., D.P. Fischer, D.E. Noakes, G.C.W. England, A. Rycroft, H. Dobson and I.M Sheldon.2007. The relationship between uterine pathogen growth density and ovarian function in the postpartum dairy cow. Theriogenology 68: 549-559.
Table- 1: Prevalence of uterine microbial flora in each stage of pregnancy of Nili-Ravi buffalo. Name of Bacteria Prevalence Total Pregnant Total Non-Pregnant Total Pregnant then No. (%) No. / % No./ % Aborted No./ % E. coli 50 (100) 34/68 12/24 4/8 S. aureus 49 (98) 34/69.38 11/22.44 4/8 Lactobacillus spp.
43/86
28/65.12
11/25.58
4/9.3
Micrococcus spp.
13/26
12/92.30
1/7.69
0/0
S. epidermidis
7/14
4/57.14
2/28.57
1/14.28
Proteus spp.
6/12
4/66.67
2/33.33
0/0
Citrobacter spp.
5/10
0/0
1/20
4/80
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Buffalo Bulletin 2013 Vol.32 (Special Issue 2): 1088-1090
Detection of bovine viral diarrhea virus prevalent in dairy herds of Punjab, Pakistan Humayun GOHARa, Masood RABBANIa*, Arfan AHMADa, Nasim AHMADb, Ali Ahmad SHEIKHa and Khushi MUHAMMADc a
University Diagnostic Laboratory (UDL). University of Veterinary & Animal Sciences, Lahore, 54000, Pakistan. b Department of Theriogenology, University of Veterinary & Animal Sciences, Lahore, 54000, Pakistan. c Department of Microbiology, University of Veterinary & Animal Sciences, Lahore, 54000, Pakistan *Corresponding email:
[email protected] ABSTRACT Bovine viral diarrhea (BVD) virus is a positive sense RNA virus having genome of approximately 12.3 kb. in length and is one of the most insidious and ubiquitous virus of bovines throughout the world. Isolation and identification of BVD virus on cell culture is labour intensive. Other diagnostic techniques used are not cost effectiveness. Therefore in this study an antigen-capture ELISA was used to detect BVDV in dairy animals at selected areas of Punjab and to evaluate comparative prevalence of the virus in cattle and buffaloes. Out of the test blood samples (n=184) originated from public (n=4) and private sector (n=3) dairy farms, 16.85% cattle and 6.31% buffalo were positive for BVD virus. Prevalence of the virus on public farms was significantly higher than that of private farms (p
