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70 cow buffalo showed 325 total heats during the period from December to April. Buffoloes have been tested for studying the spring anaestrous without the influence of pre and post partum. The cyclic status was also studied after introduction in the herd of 3 bulls (on April 16). The 33.85% of heats observed revealed levels of milk progesterone lesser than 4 ng/ml (10 days after estrous). By the end of January the incidence of acyclic heats up to 56.76% of March-April. Spring anestrous occur independently of parturition and beginning of milk production. Data showed that spring anestrous is strictly dependent to the seasonal period, to the age of the animals as well as to the interval after parturition. The restoration of sexual promiscuity bring about the disappearance of acyclic status in the 40% and 50% of cow buffaloes after respectively 1 and 3 months showing the importance of the bull effect.

La prova sperimentale ci permette di valutare positivamente l'adozione della sincronizzazione dei calori nella bovina podolica ai fini di poter effettuare l'incrocio con tori di razze italiane con attitudini sarcopoietiche più spiccate.

Data Mediterranean buffalo heats (324 spontaneous and 97 inducted by alfaprostol) have been examined. Three different levels of average temperature, thermic range, rainess and daylight were refered. Results showed that reproductive activity of buffalo breed in Italy is influenced by climatic conditions. A delay of the pituitary response to the ovaric steroisds, either in spontaneous or induced heats, occurs beyond 11 hours of daylight and with temperature below 8°C (cold springs). The oldest buffaloes feel mostly the climatic conditions, particularly when rainfall reduces.

From the end of November to the middle of April, heats duration and ovulation ways were studied on 70 buffalo cows kept apart from bulls. Resultsd referred to 324 spontaneous heats and 102 inducted by Alfaprostol. It was found a mutual relation between heats duration and ovulation time either in controls (r=0.59***). During the months characterized by a decrease of reproductive activity it is necessary to use both progesterone determination in milk and prostaglandine administration. Trough these devices a greater number of useful ovulations can be obtained in 48Hrs. In December, when the reproductive activity is more favourable, Alfaprostol administration proved to be useful considering it alowed to operate also in case of silent heats, which occur frequently (34.3% of spontaneous heats observed) during this period.

Authors have estimated the voluntary intake of a diet with 10% of crude protein on dry matter (d.m.) composed of ensiled maize stalks and full-fat soybean flakes in buffaloes (15 months of age) and rams (2.5- 4 years of age).The d.m. voluntary intake resulted higher for buffaloes (67.8 vs 41.8). The Apparent Digestibility Coefficients were almost similar for the two species. The kind of diet influenced the ruminal fluid characteristics for each species.

The domestic buffalo (Bubalus bubalis) is characterized by a worldwide distribution and its production is expressed within both intensive systems and marginal areas. Although a domesticated species since long time, over the years, the buffalo has scarcely benefitted of the strategies held in place for genetic improvement, as opposed to the cattle counterpart. Only at the beginning of the 1990s, the problems related to the enhancement of buffalo productivity through the implementation of reproductive technologies, namely the use of AI, have begun to be tackled. In this chapter, a report on the use of reproductive technologies in buffaloes will be presented, highlighting the early attempts at such and the remarkable more recent achievements in different areas, ranging from in vivo and in vitro embryo production to transgenesis, cloning, and stem cell technologies. Finally, an outlook to more pragmatic aspects will be given, such as the cost-to-benefit ratio of some of the reproductive strategies discussed.

The aim of this work was to evaluate whether the residual amount of γ-glutamyl-transferase (GGT) and alkaline phosphatase (ALP) in bovine sperm after freezing/thawing is correlated with fertility parameters, including blastocyst rates after in vitro fertilisation (IVF). The enzyme activities were determined in both spermatozoa and supernatant after centrifugation. While ALP was only correlated with sperm viability, GGT activity was correlated with sperm motility (rs = .4; p

The aim of the study was to evaluate whether resveratrol supplementation of bovine culture medium improves in vitro blastocyst development, embryo cryotolerance and cell numbers. Abattoir-derived oocytes were matured and fertilized in vitro according to standard procedure. Twenty hours after IVF, zygotes were cultured in SOF medium, supplemented with 0 (control, n=439), 0.25μM (n=422), 0.5μM (n=447) and 1μM resveratrol (n=416). On Day 7 (IVF=Day 0) blastocysts were vitrified by cryotop in 16.5% ethylene glycol, 16.5% dimethyl sulfoxide and 0.5M sucrose. Development rate, i.e. the percentage of embryos resuming development to reach a more advanced stage, and hatching rate were evaluated after 24 and 48h culture. Blastocysts cultured with (0.5μM) and without resveratrol underwent differential staining to count inner cell mass (ICM) and trophectoderm (TE) cells. Resveratrol during culture did not increase blastocyst yields (57.1, 57.7, 59.2 and 46.6%, respectively in 0, 0.25, 0.5 and 1μM resveratrol). However, 0.5μM resveratrol improved embryo cryotolerance compared to the control, as indicated by higher development rates (67.3% vs 50.3%, respectively; P

Simple Summary Congenital malformations (due to genetic causes) represent a hidden danger for animal production, above all when genetic selection is undertaken for production improvements. These malformations are responsible for economic losses either because they reduce the productivity of the farm, or because their spread in the population would decrease the total productivity of that species/breed. River buffalo is a species of increasing interest all over the world for its production abilities, as proved by the buffalo genome project and the genetic selection plans that are currently performed in different countries. The aim of this review is to provide a general view of different models of congenital malformations in buffalo and their world distribution. This would be useful either for those who performed buffalo genetic selection or for researchers in genetic diseases, which would be an advantage to their studies with respect to the knowledge of gene mutations and interactions in this species. Abstract The world buffalo population is about 168 million, and it is still growing, in India, China, Brazil, and Italy. In these countries, buffalo genetic breeding programs have been performed for many decades. The occurrence of congenital malformations has caused a slowing of the genetic progress and economic loss for the breeders, due to the death of animals, or damage to their reproductive ability or failing of milk production. Moreover, they cause animal welfare reduction because they can imply foetal dystocia and because the affected animals have a reduced fitness with little chances of survival. This review depicts, in the river buffalo (Bubalus bubalis) world population, the present status of the congenital malformations, due to genetic causes, to identify their frequency and distribution in order to develop genetic breeding plans able to improve the productive and reproductive performance, and avoid the spreading of detrimental gene variants. Congenital malformations most frequently reported in literature or signaled by breeders to the Department of Veterinary Medicine and Animal Production of the University Federico II (Naples, Italy) in river buffalo are: musculoskeletal defects (transverse hemimelia, arthrogryposis, umbilical hernia) and disorders of sexual development. In conclusion this review put in evidence that river buffalo have a great variety of malformations due to genetic causes, and TH and omphalocele are the most frequent and that several cases are still not reported, leading to an underestimation of the real weight of genetic diseases in this species.

The use of sexed semen in farm animal production and genetic improvement has been shown to be feasible with variable degree of efficiency in a number of species, and proved to be economically viable in cattle. In the last two decades, various newly developed reproductive technologies applicable in buffaloes have mushroomed. Recently, following the birth of the first buffalo calves using AI with sexed semen, commercial interest to exploit sexing of semen in this species too is aroused. In order to verify the successful adoption of this technology in the buffalo, the present study on the use of sexed semen for AI was carried out and compared with conventional artificial insemination using nonsexed semen. A total of 379 buffalo heifers were used for synchronization of ovulation using the Presynch protocol in the South of Italy. Selected animals at the time of AI were randomly allocated to three different experiment groups: (1) 102 animals subjected to AI in the body of the uterus with sexed semen (SS body); (2) 104 animals subjected to AI in the horn of the uterus with sexed semen (SS horn); and (3) 106 animals subjected to AI in the body of the uterus with conventional nonsexed semen (NSS body). Semen of three buffalo bulls was sexed by a collaborating company and commercially distributed in 0.25 mL straws with a total of 2 million sexed spermatozoa. Pregnancy rates were first assessed at Day 28 following AI, and rechecked at Day 45 by ultrasound. Pregnancy rates were nonsignificantly different between animals inseminated with sexed or nonsexed semen: 80/206 (38.8%) and 40/106 (37.7%), respectively (P = 0.85). However, site of insemination of sexed semen affected pregnancy rate significantly as higher pregnancy rates were obtained when sexed semen was deposited into the body rather than the horn of the uterus: 46/101 (45.5%) and 34/105 (32.3%), respectively (P = 0.05). In conclusion, the use of sexed semen in buffalo heifers gave satisfactory and similar pregnancy rates when compared with conventional nonsexed semen. Deposition of sexed semen into the body of the uterus, however, increased pregnancy rates significantly.

This review brings together information on ovarian physiology in buffaloes including folliculogenesis, ovulation, and the development and function of the corpus luteum. Features of embryonic development are also considered. The buffalo is classified as a short-day breeder but in equatorial zones can show oestrous cycles throughout the year provided that nutrition is adequate to maintain reproductive function. In sub-tropical zones and at higher latitudes, day length is often the major determinant of reproductive function including the occurrence of regular oestrous cycles, duration of oestrus, and the period to resumption of ovulation postpartum. Indeed, at higher latitudes buffaloes that give birth during the period of increasing day length may not show a resumption of ovulation until the following period of decreasing day length. This can have a major impact on the productive value of buffaloes and requires the development and utilisation of practical and effective assisted breeding technology for out-of-season breeding in buffaloes. Embryonic development in buffaloes occurs at a faster rate than in cattle and this has implications for the earlier establishment and functionality of the corpus luteum in buffaloes. It would appear that the interrelationships between the development of the early conceptus, corpus luteum function, uterine preparation, and maternal recognition of pregnancy, are more closely time-bound in buffaloes compared with cattle. The phase of embryonic attachment would seem to be a critical period for determining the reproductive outcome in buffaloes.

The aim of the present study was to examine whether treatment with a GnRH agonist, hCG or P(4) on Day 25 after AI increased P(4) concentrations and reduced the incidence of embryonic mortality (EM) in pregnant buffaloes mated in mid-winter in a Mediterranean environment. The trial was carried out in two farms characterized, in previous years, by low (LEM Group), 153 buffaloes (DIM=150 +/- 7 days), and high (HEM Group), 284 buffaloes (DIM=163 +/- 5 days), incidence of embryo mortality. Animals were synchronized by Ovsynch-TAI Program and artificially inseminated. On day 25, pregnant buffaloes were randomly assigned to four groups: Control (no treatment), GnRH agonist (buserelin acetate, 12.6 mu g), hCG (1500 IU) and P(4) (341 mg of P4 i.m. every 4 days for three times). Progesterone (pg/ml) was determined in milk whey on Days 10, 20 and 25 after AI in all buffaloes and in Days 30 and 45 only in buffaloes pregnant on day 25 and assigned to four groups of treatment. Pregnancy diagnosis was undertaken on Day 45 by ultrasound. All treatments increased P(4) milk whey and reduced embryonic mortality in buffalo cows bred in the farm characterized by high EM.

The percentages of dry matter (DM), protein, casein, fat, lactose, solids-not-fat and ash of milk and the characteristics of the curd (31 components and/or characteristics) were evaluated on 326 milk samples collected from 60 buffaloes every 50 days. The animals were half sib (same father or same mother ascertained by DNA test) and were bred in two farms. Relationships between the values of components and/or characteristics were also calculated. Significant relationships were found among DM percentage of milk and its components, cheese yield, curd characteristics, ratio between the studied cheese yields and protein percentage. Protein, casein and fat showed positive relationships, whereas lactose presented negative relationships. The association among proteins and real cheese yield or the dry matter of the curd that derived from 1 litre of milk (CDM), was lower than that with calculated cheese yield. This difference strongly decreased if proteins percentage, corrected for urea value, was utilized in cheese yield calculation. No relationship was found between proteins and urea in milk, but a direct relation was present between proteins and proteins adjusted for urea. This relation affects a correct estimation of cheese yield, because the percentage of true proteins in milk, corrected per urea was 4.52 vs. 4.64.

The growing rate of 240 male buffaloes fed ad libitum a diet characterized by 0.9 MFU/Kg dry matter (DM), 14% CP and 38:62 forage:concentrate ratio (Diet A) was evaluated starting from a mean age of 148 (Group 1), 218 (Group 2), 302 (Group 3), 320 (Group 4), 374 (Group 5) and 596 (Group 6) days. Data were compared at the weight of 400 kg and at the age of 550 days, values that were reached by all the subjects. The earlier administration of Diet A increased daily weight gain and reduced DM intake/die and feed conversion index compared to late administration. This trial further confirms the importance of satisfying nutritional requirements during the first months of life in buffalo species, due to the fact that these animals are not able to show a compensative weight gain like bovines.

The aim of this trial was to evaluate ghrelin response to milk administration in 20 days old buffalo calves. The trial was carried out on 5 female buffalo calves with a mean age of 21.2±2.8 days. Five blood samples were collected from each animal into EDTA tubes, starting at 07.00 until 15.00, at 2-h intervals. At 09.00, after the second blood sample, replaced milk was administered to the calves. Blood samples were immediately placed at 4°C until processing, which was performed on the same day. We used real-time reverse transcription PCR system to detect the expression of ghrelin mRNA levels in blood of buffalo calves. Two calves showed a low ghrelin concentration at the start of the trial (Group A = low ghrelin concentration) and three calves a high ghrelin concentration (Group B = high ghrelin concentration). Ghrelin expression was significantly higher either two hours (P

Several authors recorded that buffalo shows delayed first calving age and a long intercalving period. In this lecture are reported some studies which demonstrate that, if the nutritional requirements throughout growing are ensured and the culling rate is reasonable, the first calving age and the intercalving period are similar to those recorded in dairy cattle. It is worth pointing out that these conclusions come from trials performed in farms, where the out of breeding season mating technique is applied, and, hence, the fertility is affected, because mating is limited to 8 months out of 12.

This review aims to clarify some aspects of early buffalo reproduction and ovarian activity with special regard to corpus luteum (CL) activity. Puberty has been regarded as a complex event that is signalled by the first cyclic progesterone secretion at >1 ng/ml, in tight relation to other hormones such as luteinizing hormone (LH) and growth hormone (GH). CL activity has been described in relation to the peculiarities of the oestrus cycle of the species, such as seasonal reproductive aptitude and the bull effect. The importance of the CL in pregnancy establishment and maintenance has also been revised in the light of the results of the latest research about embryo mortality in buffalo.

The aims of this study were to evaluate the effects of an intravenous or intramuscular PGF2alpha analogue administration on the day of estrus on progesterone concentration and pregnancy rate in buffaloes undergoing artificial insemination (AI). To this end, two experiments were carried out. The first study was performed on 72 Mediterranean buffaloes synchronized by the Ovsynch-TAI Program. On the day of estrus only animals considered in heat were divided into four groups: Groups IVC and IMC received, respectively, an intravenous or intramuscular injection of cloprostenol (0.524 mg), whereas control Groups IVS and IMS received the same injections of saline. Milk samples were collected daily from each animal to assess progesterone concentration in the whey by RIA method. In addition on alternate days, buffaloes underwent transrectal ultrasound analysis. The second study was carried out on 385 buffaloes synchronized by the Ovsynch-TAI Program. On the day of AI, animals were divided in four groups, as described in experiment 1. Pregnancy rate was evaluated either on day 26 or day 45 and embryonic mortality rate was recorded. Statistical analysis was performed by ANOVA and chi2 test. A higher (P

Thirteen male river buffaloes, 119 females with reproductive problems (which had reached reproductive age but had failed to become pregnant in the presence of bulls) and two male co-twins underwent both clinical and cytogenetic investigation. Clinical analyses performed by veterinary practitioners revealed normal body conformation and external genitalia for most females. However, some subjects showed some slight male traits such as large base horn circumference, prominent withers and tight pelvis. Rectal palpation revealed damage to internal sex adducts varying between atrophy of Mullerian ducts to complete lack of internal sex adducts (with closed vagina). All bulls had normal karyotypes at high resolution banding, while 25 animals (23 females and 2 male co-twins) (20.7%) with reproductive problems were found to carry the following sex chromosome abnormalities: X monosomy (2 females); X trisomy (1 female); sex reversal syndrome (2 females); and free-martinism (18 females and 2 males). All female carriers were sterile. Copyright (C) 2008 S. Karger AG, Basel.

The aim of this study was to evaluate the length and the characteristics of the oestrous cycle in Italian Mediterranean buffalo cows, undergone synchronization of ovulation. The trial was performed on 32 buffaloes synchronized by the Ovsynch Program, which consists of an injection of GnRH on day 0, PGF2α on day 7 and GnRH on day 9. Starting on day 10 (Day 0 of the new cycle). Buffaloes undergone ultrasound examination of the ovaries on alternate days until the following heat. Follicular growth and corpus luteum formation and dimensions were recorded as well as the number of follicular waves. Statistical analysis was performed by ANOVA. Four animals (12.5%) did not show signs of oestrous and were excluded from the trial. The mean length of the oestrous cycle was 23.7±3.4 days. In particular, 1 animal (3.6%) showed an oestrous cycle characterized by 1 follicular wave with a length of 16 days, 17 subjects (60.7%) showed 2 follicular waves with a cycle length of 22.4±2.3 days and 10 buffaloes (35.7%) showed 3 follicular waves with a cycle of 26.8±2.0 days. These results confirm previous reports performed in buffalo species, although the cycle resulted longer in the 3-waves group.

Here reported are data relative to the chemical composition/characteristics of the milk, of milk samples collected every 50 days from 60 buffaloes (326 samples) that were half-sibs (through paternal or maternal lineage, ascertained by DNA test), bred in two farms (S.A. and M respectively n. = 27 and n = 33). The subjects were divided in relation to the productive level (> vs. < 2601 kg/lactation). The production level did not influence the theoretical yield (ThCY), the yield at 4 h (FCY) and at 28 h (28CY), the curd dry matter (CDM) and the ratios between ThCY and real yields (FCY, 28CY and CDM). Compared to the less productive subjects, despite similar percentage of proteins, the more productive subjects had a higher percentage of casein (3.73 vs. 3.64; P < 0.05), a greater incidence of the latter on total protein (80.52 vs. 79.60; P < 0.05), higher content in lactose (4.88 vs. 4.81; P < 0.01) and urea (42.44 vs. 40.48; P < 0.05) and a lower BCS value (6.79 vs. 7.25; P < 0.05). These differences are not always confirmed in both farms. Regardless of the production level, subjects of farm S.A. had a lower production/lactation (2524 vs. 2817 kg; P < 0.01), higher values of R and k20 (P < 0.01), lower value of a30, a higher SH value and lower contents in urea and solid not fat. They also showed a lower ThCY yield and a lower ratio between ThCY and 28CY when proteins were corrected for the urea content, and a higher ratio between 28CY and the percentage of proteins, both when corrected or not for the urea content. It is difficult to explain the influence of the production level on the casein content whereas the greater lactose content in more productive buffaloes may be accounted for by the greater persistence of lactation curve. The higher urea content found in buffaloes with a greater galactopoietic attitude may be due to a greater DM intake caused by the inadequacy of the diet, hypothesis confirmed by the lower BCS observed in the more productive subjects.

The aim of this study was to verify the ratio between real cheese yield at 28 hours and proteins, after values correction according to the regression analysis. Milk characteristics of subjects characterized by a 28CY/protein ratio lower (Group A; n=212) or higher (Group B; n=108) than 56 were analysed by ANOVA in order to evaluate differences between the following milk characteristics: the lactodinamographic parameters (Formagraph, Foss, DN); the complete physic-chemical composition; fat (F), protein (P), casein, lactose, ash, urea, pH and SH; the somatic cells content (SCC) using (Milkoskan); the total aerobic mesophilic flora (TAMF) by the dilution method. Chemical composition of the curd was also determined (protein, fat and ash; ASPA, 1995). The milk protein content was adjusted for the non proteic N content determined in milk as urea (corrected P); the theoretic cheese yield (ThCY) was obtained by the following formula: cheese yield =milk x(-0.88 + 3.50 x P(%)+1.23 x F(%)) x 100 -1 (Altiero et al., 1989) and the ratio between ThCY/FCY and ThCY/28CY were calculated. Other calculated variables were: FCY/Corrected P, 28CY/Corrected P, CDM/Corrected P. Regression analysis was carried out between real cheese yield at 28 hours/proteins ratio and 28CY vs. all the parameters resulted different between the two groups of buffaloes. Real cheese yield at 28 hours/proteins ratio and 28CY were corrected and ANOVA was repeated on corrected data, in order to verify the modifications of the values. These correction reduced but did not eliminate the differences.

The aim of this study was to evaluate the relationships between 44 components and/or characteristics of milk samples collected every 50 days from 60 buffaloes (326 samples and 14,344 values). The animals were half sib (same father or same mother) and were bred in two farms. Significant associations but with a low values of coefficient of correlation were present, demonstrating that lactodinamographic parameters marginally affect cheese yield.