Allometry of Organs and the Gastrintestinal Tract from Water Buffaloes (Bubalus bubalis) Finished in Feedlot

The objective of present study was to evaluate the relative growth of organs and viscera from water buffaloes. Fifteen Mediterranean intact males, averaging, 356.7 kg initial live weight and twenty four months of age, were used. The animals were ramdomly assigned into three groups (categories). One group was ramdomly assigned to immediate slaughter (AR), the rest two groups were full-fed a ration containing 50% concentrate, dry matter basis until reaching the slaughter weights of 450 and 500kg, respectively. At slaughter the empty body weight was determined and the weights of head, feet, leather, lungs, heart, liver, spleen, rumen-reticulum, omasum, abomasum, small intestine, large intestine were recorded. Regression equations of log weight of organs and viscera as a function of log empty-body-weight (EBW), were fitted. All body components studied, with exception of liver and spleen, developed slower than in relation to EBW.

Using Slaughter Weight to Predict Weight and Yield of Primal Cuts of Carcass from Buffaloes

The knowledge of the meat production from different buffalo breeds and their crossings in different feeding systems becomes necessary for the supply of subsidies to whole productive meat chain. Some quantitative carcass traits of Mediterranean buffaloes bulls, finished in feedlot, with initial age of fourteen months and 330 kg live weight, slaughtered with 450, 480, 510 and 540 kg, were evaluated. The diet contained 13% crude protein, 2.68 Mcal digestible energy/kg DM and a roughage : concentrate ratio of 25:75. Regression equations for prediction weight and yield of primal cuts of carcass as a function of slaughter weight were obtained. Carcass dressing percent increased as the slaughter weight increased (49.2; 49.5; 49.7; and 49.9%). The Pistola Style cut weight although increasing linearly in weight (108.2; 117.6; 124.0 and 130.7 kg) as the slaughter weight increased, declined linearly when expressed in relation to cold carcass weight (49.5; 49.0; 48.6 and 48.2%). In this experimental conditions Mediterranean young bulls slaughtered between 450 to 540 kg of live weight showed increasing yields of cold carcass, forequarter and thin flank.

Body Composition and Net Energy Requirements for Maintenance of Non-Castrated Water Buffaloes (Bubalus bubalis)

The research aimed to estimate body contents of protein and energy and net requirements of energy for maintenance of buffaloes, slaughtered at different stages of maturity. There were used 14 Mediterranean intact males with initial average body weight of 352.2 +/- 24.3 kg and average age of 24 months. The animais were randomly divided into four experimental groups. One group was designed to slaughter at the beginning of the experimental period (IS). The animals of another group were restricting fed, receiving, individually, levels of protein and energy 15% above maintenance (RF). The animals of the two remaining groups were individually fed ad libitum (SW450 and SW500) to reach weights corresponding to 100 and 110 percent of the mature weight of the buffalo cows (respectively 450 and 550 kg). The ration contained ground-corn cobs, soybean meal, urea, minerals, and signal-grass (Brachiaria decumbens) hay, with a concentrate: roughage ratio of 50: 50 and 13% of crude protein on a dry matter basis. To estimate changes in body composition inside the range of weights included in the trial, linear regression equations of log protein (kg), fat (kg) and energy (Mcal) as a function of log empty-body-weight (EBW), in kg, were fitted. Energy requirements for maintenance were obtained as estimated heat production at zero level of energy intake. Buffaloes submitted to fattening in feedlot presented early body fat deposition, and had with the same live weight lower protein content and higher fat content and energy per unit weight than european-zebu crossbred cattle.

Bone Marrow'S Harvest in The Coxal Tuberosity for Isolation and Culture of Mesenchymal Stem Cells of Buffaloes (Bubalus Bubalis)

Several studies with mesenchymal stem cells (MSCs) have been developed in many species because of its ability to differentiate into other mesoderm lineages, capacity of self-regeneration, low immunogenicity, paracrine, anti-inflamatory, immunomodulatory and antiapoptotic effects which make then a promissory source to be used in therapeutic strategies. The aim of this study is to report the technique of harvest of bone marrow (BM) in the coxal tuberosity (CT) of buffaloes. For this, the animals were selected, identified and contained in a stock. Then trichotomy was performed in the region corresponding to the CT. After identifying the anatomic site it was performed antisepsis, local anesthetic block and introduction of a myelogram's needle (Lang(R)) for BM aspiration. Once the needle was firmly fixed in the CT, the mandril was removed and proceeded to BM aspiration with a syringe (20 mL) containing 1 ml of heparin at 1000 IU / mL and 1 mL of PBS. After the collection, each sample collected was manually homogenized, identified and referred to the LRACT - FMVZ / UNESP-BRAZIL for the correct processing. The anatomical site tested showed to be an alternative site of harvest of BM once provided the appropriate isolation and culture of the mononuclear fraction. Moreover, the procedure was performed without difficulty and with great security. Based on this, it can be conclude that CT is an excellent anatomical site for isolation and culture of MSCs and the proposed technique is viable and feasible to be held in buffaloes.

Isolation, Culture and Differentiation of Buffaloes Bone Marrow Mesenchymal Stem Cells Obtained from the Coxal Tuberosity

Currently, much attention has been devoted to the renewal of knowledge about Stem Cells and Cell Therapy in domestic species. In this sense, the present work aimed to develop a methodology for collecting, processing and cultivation of mesenchymal stem cells obtained from bone marrow of coxal tuberosity in buffaloes. The collection was performed using a Komiyashiki needle, which was introduced in the coxal tuberosity and the bone marrow aspirated into a heparinized syringe with the aid of negative pressure. Directly after collection samples were processed at the laboratory at FMVZ - UNESP. The samples took approximately 32 days to reach 80% confluence, when the first passage and differentiation was performed. To confirm the mesenchymal origin, cells were induced to differentiate into adipogenic and osteogenic lineages. Samples showed morphological changes during differentiation protocol, but not all presented production of extracellular deposits of calcium or intracellular fat droplets, observed after staining with Alizarin Red and Oil Red respectively. Compared with the material obtained from other species and processed in the same laboratory, the primary culture was longer. Therefore, more studies are needed to standardize the age of animals used and to test other inducers of cell differentiation.

Effect of testicular thermoregulation on the quality of buffalo sperm

The process of spermatic division and differentiation (spermatogenesis) occurs with intratesticular temperature lower that the corporal temperature and for that is essential that the testicular thermoregulation mechanism occurs properly. For evaluation of the scrotal surface temperature can be used the infrared thermography or testicular sensors, besides that, can be evaluated the blood flux in the spermatic cord through the Doppler ultrasonography. Thus, the aim of this study is to analyze the testicular thermoregulation in adult buffaloes through scrotal thermography and Doppler ultrasound of testicular artery and verify its effect on sperm quality. For that were used seven healthy buffaloes, with age of 3 and 4 years, of the Murrah breed. The animals were subjected to 3 semen collections using artificial vagina, with one day of interval. In addiction, the retal temperature measurement (RT) with dry bulb thermometer, the measurement of scrotal surface temperature (SST) and body surface temperature (BST) through infrared thermography and the pulsatility (PI) and resistivity (RI) index of testicular artery by Doppler ultrasonography, were performed using 2 distinct moments: animals previously placed to shade (M1) and animals subjected to 4 hours of sun (M2). All parameters were compared by T test and the correlations were performed by Pearson test using the In Stat Graph Pad 3 (R) program. The significant level considered was 5%. There was an increase (p<0,05) of RT, SST, SNT and RI in M2. increasing trend was observed (0,05>p>0,01) PI and RI between M1 and M2. There was a low correlation between SST and semen quality. The results of this study allow us to conclude that adult buffaloes have low ability to perform body and testicular thermoregulation in situations of enviromental heat stress. However, this low capacity of testicular temperature maintenance demonstrated no correlation with the sperm kinetic parameters and sperm morphological defects in buffalo spermatozoa.

Morphometric and Functional Characteristics of Buffaloes and Cows Corpus Luteum at Different Stages of the Estrous Cycle

The aims of the present study were to evaluate the morphometry of corpus luteum (CL) and progesterone (P-4) plasma concentration of 86 buffaloes (33 pregnant and 53 non-pregnant) and 95 cows (36 pregnant and 59 non-pregnant) at the moment of slaughter. Seventy CLs of buffaloes and 110 CL of cattle were analyzed. The CL classified as II and III were more common in both species (35.7 and 41.4% for buffaloes and 43.6 and 35.5% for cows). The 29 non-pregnant buffaloes had a total of 36 CL, being 19.4% CLI; 33.3% CL II; 27.8% CL III and 19.4% CL IV. The 51 nonpregnant cows had a total of 71 CL, being 26.8% CL I; 47.9% CL II; 21.1% CL III and 4.2% CL IV. The average diameters of bubaline and bovine CL were 5.2 +/- 0.9 and 6.4 +/- 1.8 mm (CL I); 17.6 +/- 2.6 and 19.8 +/- 3.2 mm (CL II); 17.2 +/- 2.1 and 20.0 +/- 3.2 mm (CL III); 7.8 +/- 1.8 and 8.7 +/- 2.7 mm (CL IV), respectively. The mean plasma concentrations of P4 were 5.6 (CL I); 5.4 (CL II); 4.7 (CL III) and 0.5 (CL IV) ng/mL for buffaloes and 0.02 (CL I); 6.3 (CL II) and 6.4 (CL III) ng/mL for cows. In both species, P4 concentration was similar between stages II and III. The results indicated that the characterization of the CL provides important information about the status of estrous cycle.

Values of 13,14-Dihydro-15-Keto-PGF(2 alpha), Progesterone and Oestradiol After Injection of Prostaglandin at Different Periods of Buffalo Puerperium

Knowledge of the effectiveness of prostaglandins in uterine involution process led to the development of protocols with its analogues in postpartum period. However, this hormone mechanism of action is not yet fully elucidated. Thus, the objective of this study was to verify if chloprostenol administration, at early or intermediary puerperium, can induce changes on progesterone, PGFM and oestradiol plasma concentrations. 30 Murrah postpartum buffaloes were randomly divided into three groups: CONT (saline, n = 10); CLO2 (chloprostenol at days 2 and 5 postpartum, n = 10) and; CLO15 (chloprostenol at days 15 and 20 postpartum, n = 10). Blood samples were collected from jugular vein to measure progesterone, PGFM and oestradiol plasma concentrations at days 2, 7, 14, 21 and 28 postpartum. CLO2 group presented lower progesterone and PGFM plasma concentrations in relation to CONT and CLO15 groups (0.23 +/- 0.00 and 0.32 +/- 0.11, 0.19 +/- 0.00 and 0.23 +/- 0.11, 0.23 +/- 0.00 and 0.30 +/- 0.19, for groups CONT, CLO2 and CLO15, respectively; P < 0.05). There was no significant difference in oestradiol plasma concentration between experimental groups (P > 0.05). Prostaglandin synthetic analogue administration induced hormonal changes in postpartum buffaloes, which can partially explain its positive effect under reproductive function of this specie.

Uterine Involution, Fluid Accumulation and Ovarian Activity after Injection of Prostaglandin at Different Periods of Buffalo Puerperium

This study aimed to evaluate the effect of chloprostenol administration, at early or intermediary puerperium, under uterine involution, intrauterine fluid accumulation and ovarian activity return. 30 Murrah postpartum buffaloes were randomly divided into three groups: CONT (saline, n = 10); CLO2 (chloprostenol at days 2 and 5 postpartum, n = 10) and; CLO15 (chloprostenol at days 15 and 20 postpartum, n = 10). Gynecological exams were performed at days 2, 7, 14, 21 and 28 postpartum, when uterine involution degree (1 to 3 scale, by transrectal palpation), intrauterine fluid accumulation (0 to 3 scale, by ultrasound exam) and ovarian activity (B-mode ultrasound exam) were evaluated. CLO2 group presented higher uterine involution (2.00 +/- 0.23, 1.66 +/- 0.23, 1.58 +/- 0.23 for groups CLO2, CONT and CLO15, respectively) and faster ovarian activity return in relation to groups CONT and CLO15 (P < 0.05). Groups CLO2 and CLO15 showed lower intrauterine fluid accumulation compared to CONT group (2.04 +/- 0.20, 1.58 +/- 0.20, 1.92 +/- 0.20 for groups CONT, CLO2 and CLO15, respectively; P < 0.05). Prostaglandin analogue administration in postpartum buffalo benefited uterine involution, lochia expulsion and ovarian activity return, improving reproductive efficiency in this specie.

Inbreeding, Average Relatedness Coefficient and Effective Population Size In Jaffarabadi Buffaloes Raised In Brazil

The major aim of this study was to evaluate the inbreeding (F), average relatedness coefficient (AR) and effective population size (Ne) in the Jaffarabadi buffalo breed from Brazil. Pedigree information of 1,272 animals born from 1966 was used. The effective population size was calculated in two ways: first, computed via individual increase in inbreeding and second estimated by individual increase in coancestry. The known generation numbers were 1.24, 1.76 and 2.64 for complete, equivalent and maximum generation, respectively. The effective size computed via individual increase in coancestry was small with a value of 10.82 +/- 1.29. The effective size computed by individual increase in inbreeding (10.40 +/- 3.69) was very similar but a little smaller than the previous reported value. The average values of F and AR for the population reference (1,059) were 4.22 and 12.5 percent. The mean of F for inbred animals (319) was 14.0%. The F and AR means were 5.7 and 13.3% for animals with at least 1.5 known equivalent generation and 9.3 and 15.97% for individuals having at least 2.5 equivalent generations known. It was found 78 matings between half sibs (6.14%) and 67 matings (5.27%) between parent-offspring. The estimated inbreeding increase per generation by considering maximum generation, complete generation and equivalent generation were 1.21%, 5.18% and 3.57%, respectively. Considering the uncompleted pedigree, the estimated inbreeding for the reference population could be underestimated.

Reference Gene Selection for Gene Expression Analysis of Oocytes Collected from Dairy Cattle and Buffaloes during Winter and Summer

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Progestin-Based Protocol for Synchronization of Follicular Wave Emergence in Buffaloes During Summer and Winter

The aim of the present study was to evaluate the effects of type of norgestomet auricular implant (new - N or previously used during 5 days - U), season of the year (summer - S and winter - W), and parity (12 heifers - H and 23 cows - C) on synchronization of follicular wave emergence in buffaloes. For this purpose, 35 buffaloes were examined daily by ultrasonography until follicular wave emergence was detected. Data were analysed by ANOVA, using PROC GLIMMIX. No interactions were observed in none variables. Time of follicular wave emergence and number of follicles at emergence were not affected by type of implant or season of the year. Parity also did not influence the number of follicles at emergence. However, follicular wave emergence occurred later in heifers than in cows. In conclusion, the previous use of a norgestomet auricular implant independent of the season of the year does not affect the time or the number of follicles at follicular wave emergence in buffaloes. Nevertheless, although heifers and cows had a similar number of follicles at emergence, the time of follicular wave emergence occurs earlier in cows than in heifers.

Efficiency of OPU-IVEP-ET of Fresh and Vitrified Embryos in Buffaloes

The present study aims to report ovum pickup (OPU), in vitro embryo production (IVEP) and embryo transfer (ET) outcomes of fresh and vitrified buffalo embryos. For this purpose, 36 buffalo donors were submitted to 11 OPU sessions (n = 201). A total of 998 oocytes (5.0 +/- 0.5/donor/session) and 584 viable oocytes (2.9 +/- 0.3/donor/session) were recovered. Viable oocytes (grades 1, 2 and 3) were subjected to IVM, IVF (D0) and IVC. On D2, 54.5% of cleavage rate was obtained. Embryo yield on D7 was 44.9% (grade 1: 229 embryos, grade 2: 5 embryos and grade 3: 28 embryos). From this total, 115 fresh (grades 1 to 3) and 70 vitrified embryos (only grade 1) were transferred into recipients previously synchronized with fixed time embryo transfer (FTET) protocol. Vitrification was performed using the cryotop method. Pregnancy diagnosis in fresh and in vitrified groups were, respectively: 43.5% (50/115) and 37.1% (26/70) on 30 days after embryo transfer, and 41.7% (48/115) and 31.4% (22/70) on 60 days after embryo transfer. In conclusion, our results demonstrate the possibilities for commercial use of the techniques of OPU, IVEP and ET of fresh and vitrified embryos in buffaloes.

Pregnancy Monitoring of In Vitro Produced Embryos in Buffaloes

In the present study, pregnancies obtained from 115 in vitro produced embryos were monitored by ultrasonography on days 30 and 60 after embryo transfer (ET), and at calving. Additionally, the health of newborns and recipients were also evaluated. On day 30 after ET, positive pregnancy was diagnosed in 50 animals (43.5%). A total of 8 fetal mortalities (16.0%) were verified from 30 days until calving, in which 2 occurred from 30 to 60 days after ET (4.0%), and 6 occurred from 60 days until calving (12.0%). In this last period of pregnancy, 3 pregnancy losses were due to abortion, and the other 3 were stillbirth. One additional animal was eliminated from the study, remaining 41 pregnancies. From these 41 pregnancies, a total of 20 female calves (48.8%) and 21 male calves (51.2%) were born. Pregnancies from female and male calves had a mean length of 309.8 and 310.9 days, respectively (range 300 to 328 days, and 297 to 320 days, respectively). Weight at calving was a mean of 31.4 and 33.8 kg for female and male calves, respectively. All calving occurred without intervention and dystocia was not observed in any case. No large offspring syndrome, hydramnios, hydroallantois, or umbilical cord anomalies were observed in calves. Delivery was normal in all recipients, and no puerperal infections, or retained placenta occurred. Suckling assistance was not necessary in any newborn. All genetic pedigree was confirmed later by DNA tests.