Follicular dynamics and oestrous detection in Thai postpartum swamp buffaloes (Bubalus bubalis).

This study characterized follicular activity and oestrous behaviour from 5 to 9 days post-calving up to the 4th ovulation postpartum (pp) in 16 multiparous (range 2-7 parities) Thai swamp buffalo cows (Bubalus bubalis), aged 4-12 years and weighing from 432 to 676 kg. Ovarian follicular activity was examined by transrectal ultrasonography (TUS) every morning. Oestrous detection was performed twice daily by direct personal observation of behaviour and for presence of clear cervical mucus discharge and indirectly by video camera recording during 21 h/day. A follicular wave-like pattern was present before the 1st ovulation leading to short oestrous cycles. Growth rates and maximum diameters of the ovulatory follicles did not differ between the 1st and 4th ovulations. However, growth rate for non-ovulatory dominant follicles (DF) before the 1st ovulation was lower than for the ovulatory follicle (p<0.05). In addition, the diameter of all ovulatory follicles (14.3 ± 0.46 mm, n=39) was significantly larger (p < 0.01) than those of the preceding last but one non-ovulatory DF (10.8 ± 0.20 mm, n = 5), but similar to the last preceding non-ovulatory DF diameter (12.92 ± 0.96 mm, n = 14). Short oestrous cycles were most common between the 1st and 2nd ovulations (93.75%, 15/16 cows, 10.2 ± 0.38 days) decreasing in prevalence thereafter (50%, 3/6 buffaloes, 12.0 ± 1.53 days). Oestrous signs were relatively vague around the 1st ovulation pp to become more easily detectable thereafter. This study suggests that properly fed swamp buffaloes could be mated successfully within 2 months pp, at their 2nd spontaneous ovulation, provided oestrous detection is at least performed daily at 06:00-08:00 hour.

Maturation competence of swamp buffalo oocytes obtained by ovum pick-up and from slaughterhouse ovaries.

This study was designed with the final goal of improving in vitro embryo production in the Thai swamp buffalo (Bubalus bubalis carabensis). Oocytes were collected by ovum pick-up (OPU) from six non-lactating multiparous swamp buffalo twice per week for 10 consecutive sessions followed by once-weekly collection for 10 consecutive sessions without hormone stimulation. In addition, oocytes were collected from slaughterhouse ovaries that were classified as follows: ovaries from non-pregnant cows with a visible corpus luteum (NPCL); pregnant cows with a corpus luteum (P); and non-pregnant cows without a corpus luteum (NP). Follicles in each group of ovaries were categorized as small (2-4 mm), medium-sized (5-8 mm) or large follicles (≥ 9 mm). The quality of the oocytes was assessed by their capacity to undergo in vitro maturation. The total number of observed follicles per session (all sizes combined) was larger in the once-weekly OPU group compared with the twice-weekly OPU group. In particular, the numbers of small and large follicles were higher in the once-weekly OPU group (5.2 ± 0.7 and 0.9 ± 0.2, respectively) than in the twice-weekly OPU group (3.9 ± 0.5 and 0.5 ± 0.1). The number of medium-sized follicles did not differ between the groups. The percentages of oocytes with an abnormal spindle morphology were not different between oocytes from the twice-weekly (30.0%) and the once-weekly (28.6%) OPU groups. A higher percentage of oocytes obtained in vitro (49.5%) exhibited nuclear abnormalities compared with those obtained in vivo (≤34.8%) after in vitro maturation. In conclusion, oocytes can be successfully collected by OPU in the swamp buffalo, without hormonal pretreatment, and per week more good-quality oocytes can be collected by twice-weekly OPU. In addition, oocytes collected from slaughterhouse ovaries can be used with the reproductive status of the cow having no influence on the maturation competence of oocytes.

Y-chromosomal variation confirms independent domestications of swamp and river buffalo.

Y-chromosomal variation in the water buffalo was analysed by sequencing of DBY, ZFY and SRY gene segments. A clear separation of the paternal lineages of the river and swamp types parallels the differences between their maternal lineages and nuclear DNA. Sequence divergence was found to be comparable to the divergence of taurine cattle and zebu, and this divergence predated domestication, confirming that river and swamp buffalo originated from different wild populations. Within a sample of 23 Thai swamp buffaloes, we identified four haplotypes with different geographical distributions, two of which were shared by Thai wild buffaloes.

Effects of thawing temperature and post-thaw dilution on the quality of cat spermatozoa.

The present study aimed to compare cat sperm quality after thawing using two different temperatures (37 and 70 degrees C) and to investigate the effects of post-thaw dilution on the sperm quality and longevity of ejaculated cat spermatozoa. Six ejaculates of each of six male cats were collected using an electroejaculator (total 36 ejaculates). The semen was frozen in 0.25-ml straws using a Tris egg yolk extender containing Equex STM paste. Four straws prepared from each ejaculate were thawed at four different occasions; (i) at 37 degrees C for 15 s, (ii) at 37 degrees C for 15 s and diluted 1 : 2 with Tris buffer (v/v), (iii) at 70 degrees C for 6 s, (iv) at 70 degrees C for 6 s and diluted 1 : 2 with Tris buffer (v/v). The percentages of motile spermatozoa, the scores of progressive motility, the percentages of spermatozoa with intact plasma membrane (using SYBR-14/EthD-1 stains) and intact acrosome (using fluorescein isothiocyanate conjugated peanut agglutinin/propidium iodide stains) were evaluated in fresh semen at 0, 2, 4 and 6 h after thawing. The thawing temperature had no effect on any sperm parameters throughout the incubation period (p > 0.05). The dilution after thawing improved sperm motility, progressive motility and acrosome integrity (p < 0.05). The thawing of cat spermatozoa and subsequently diluting with Tris buffer resulted in an immediate (at 0 h) overall (combined over temperature) percentage of motile sperm of 64.8 +/- 10.7 (mean +/- SD), a score of progressive motility of 4.0 +/- 0.5, a percentage of spermatozoa with intact plasma membrane of 64.4 +/- 12.1 and intact acrosome of 44.8 +/- 20.2. In conclusion, frozen cat semen can be thawed either at 37 or 70 degrees C and post-thaw dilution is recommended to reduce the toxic effect of some ingredients in the extender during post-thaw incubation.

Seronegative conversion in four Neospora caninum-infected cows, with a low rate of transplacental transmission.

Four Neospora-seropositive pregnant cows (prebreeding indirect fluorescent antibody (IFA) titers between 1:400 and 1:1600) were confined and observed until parturition. All cows gave birth to normal calves. Selected tissues were tested for NC by histopathology, immunohistochemical (IHC) and polymerase chain reaction (PCR). Parasite isolation was attempted in vero cell cultures. At parturition, all cows were seronegative at 1:200 and two of four cows had a titer of 1:100 when further tested. Three of four calves were not infected, as determined by negative results of precolostral serology (1:25 cut-off), histopathology, IHC and PCR. One calf was congenitally infected, as shown by the presence of a thick-walled cyst labelled by IHC in its brain, positive PCR of brain and a precolostral IFA titer of 1:100. It was concluded that NC antibody titers may drop or convert to seronegative status in chronically infected cows by the time of parturition and this finding in four of four cows indicates that this could be a common occurrence. Similarly, the finding of an infected calf with a low antibody titer indicates that precolostral serology may not be a fool-proof means of identifying calves with congenital Neospora caninum infections. These findings call into question conclusions of other studies that have estimated rates of congenital transmission of this parasite based on serological tests at calving. This study is the first confirmed report of congenital NC infection in a calf in Thailand.

Morphology and life cycle of Schistosoma sinensium Pao, 1959, from northwest Thailand.

Schistosoma sinensium Pao, 1959, was first isolated from an unidentified snail in Szechuan Province, China, and was described on the basis of adult worms and eggs recovered from experimentally infected mice. We discovered snails, Tricula bollingi Davis, and rodents, Rattus rattus (L.) and Crocidura attenuata Milne-Edwards, infected with S. sinensium near Fang in northwest Thailand. Adult worms were recovered from small veins immediately adjacent to the small intestine. Eggs first appeared in the feces of experimentally infected mice at 25 days postinfection. Cercariae first emerged from experimentally infected T. bollingi at 40 days postinfection. Shortly after emerging, cercariae were found at the water surface, their bodies flat against the meniscus and tails hanging free in the water column. Our morphometric results for adults and eggs from naturally infected hosts agree well with those presented in the original description. The occurrence of a lateral spine on the egg of this Asian schistosome has created some confusion as to the affinities of S. sinensium. A comparison of 8 character states, including snail hosts, cercarial behavior, geographical distribution, and morphology of life cycle stages, indicates that S. sinensium is more closely related to S. japonicum Katsurada than it is to S. mansoni Sambon.

Ovarian responses and oocyte recovery in prepubertal swamp buffalo (Bubalus bubalis) calves after FSH or PMSG treatment.

Stimulation of follicular growth was examined using two different gonadotropin treatments in 10 prepubertal swamp buffalo calves (8 to 12 mo old). Each calf received an ear implant consisting of 3 mg norgestromet and 5 mg estradiol valerate during hormonal treatment. Five calves were additionally administered FSH (24 mg, im) and, 2 mo later, PMSG (3,000 IU). The remaining 5 calves were first treated with PMSG followed by FSH. Ovarian responses to treatments were examined by laparotomy, 72 h after ear implant removal, and by the number of follicles (diameter > or = 0.8 cm) and corpora hemorrhagica present. Ovaries had more significant response to FSH than PMSG treatment (13.9+/-8.6 vs 5.9+/-3.3 follicles; P<0.01). Although the recovery rate tended to be lower for FSH treated (64%) than PMSG-treated (82%) animals, more oocytes/animal were harvested in the PMSG treatment (8.3+/-5.0 vs 4.6+/-3.2, respectively). The immature oocytes (n = 38) were cultured for 24 to 25 h in maturation medium (TCM-199 NaHCO3+10% fetal calf serum [FCS] in 5%CO2 in air at 39 degrees C). Oocyte maturation was assessed after fixation and staining with aceto orcein. The in vitro maturation rate was 52.6% (20/38). This study shows the possibility of harvesting oocytes from prepubertal swamp buffalo calves and maturing the oocyte in vitro.

Early embryonic development in Thai swamp buffalo (Bubalus bubalis ).

A total of 33 nonsurgical embryo collections was carried out to investigate early embryo development in Thai swamp buffalo. Collections were performed on Days 5.5, 6.0, 6.5, 7.0 and 7.5. The different stages of embryo development on these days were the 16-cell stage, compact morula, blastocyst, hatched blastocyst and hatched expanding blastocyst, respectively. In addition, some degenerating embryos and unfertilized ova were also recovered. A higher recovery rate was obtained with single embryo collection after natural estrus than after induced estrus or superovulation, 78% (7 9 ) vs 46% (6 13 ) vs 54.5% (6 11 ), respectively. A higher percentage of normal embryos was also obtained with single embryo collection after either natural or induced estrus than after superovulation, 71% (5 7 ), 83% (5 6 ) and 38% (6 16 ), respectively.

Circadian variations of plasma LH and testosterone in adult swamp buffalo bulls.

Three swamp buffalo bulls aged 1.5, 1.10 and 2 years were submitted to frequent blood sampling every 15 m during a period of 25 h using an indwelling infusion set. Plasma LH and testosterone were quantified by radioimmunoassay technique. The levels of the two hormones in each individual exhibited episodic and nonrhythmic patterns. The number of LH peaks varied according to individual, ranging from no peak in one bull to 2 in the other two bulls. The mean LH concentrations during the period of study for each bull were 0.74, 0.33 and 1.17 ng/ml. Whereas the number of testosterone peaks varied between 1-10 and the average testosterone concentrations were 0.1, 0.33 and 0.55 ng/ml for the younger to the older bulls respectively. The testosterone peaks related to the LH peaks in each individual bull.

Survival of beta and gamma races of Neotricula aperta snail intermediate hosts of Schistosoma mekongi in raw waters.

Beta and gamma races of Neotricula aperta were exposed to naturally raw waters of the Sirindhorn reservoir, Mun River and Mekong River in laboratory conditions and non supplement on food. Snails of two races could withstand well in variable qualities of water within 48 hours. The survival rates of N. aperta in three sources of raw water were 100% for the beta race and 97 to 100% for the gamma race.

Distribution of Pila polita in a southern province of Thailand.

This field study investigated the distribution of the freshwater snails of the family Ampullariidae in 18 provinces of Thailand. The introduced Pomacea canaliculata has wide distribution in the studied provinces. Pila angelica, P. ampullacea and P. pesmei still have limited distribution. Pila polita, which is absent in the south, now can be found in Phangnga, a southern province of Thailand. Moreover, P. polita is currently used as a traditional medicine of the local people for the treatment of a skin disease.

Investigation into the use of prostaglandin F2 alpha (PGF2 alpha) and oxytocin for the induction of farrowing.

This experiment was conducted in order to compare the effects of injecting PGF2 alpha alone, PGF2 alpha with oxytocin and placebo on the induction of farrowing in swine and to compare the relative effects of 3 different dosages of oxytocin (10, 20 and 30 iu per animal) when combined with PGF2 alpha (10 mg). The findings revealed that animals treated with 30 iu oxytocin farrowed within 10.6 h which was similar to those receiving PGF2 alpha only (9.4 h), but shorter than control animals (53.6 h). Animals receiving 20 and 10 iu of oxytocin farrowed within 1.4 and 1.7 h, respectively. Difficult farrowings requiring manual assistance occurred in 30%, 30%, 50% and 10% of sows given 30 iu, 20 iu and 10 iu of oxytocin and in the control group, respectively. Thirteen of 73 sows treated with PGF2 alpha farrowed within 12.6 +/- 5.3 h. Stillbirths were highest (10.2%) in the control animals whilst in the others it was under 7%. Oxytocin at dosages of 20 and 10 iu, seemed most promising in terms of synchronising farrowing following PGF2 alpha treatment in swine. However, farrowing complications were more common in these groups.

Improved cryopreservability of stallion sperm using a sorbitol-based freezing extender.

Cryopreservation of stallion semen is often associated with poor post-thaw sperm quality. Sugars are among the important components of a freezing extender and act as non-permeating cryoprotectants. This study aimed to compare the quality of stallion sperm frozen with glucose, fructose or sorbitol-containing freezing extenders. Semen was collected from six stallions of proven fertility and cryopreserved using a freezing extender containing different types of monosaccharide sugars (glucose, fructose or sorbitol). After thawing, the semen was examined for sperm motility, viability, acrosome integrity, plasma membrane functionality and sperm longevity. The fertility of semen frozen in the presence of sorbitol was also tested by artificial insemination. Sperm quality was significantly decreased following freezing and thawing (P < 0.05). Fructose was inferior for protecting sperm during cryopreservation when compared to sorbitol and glucose (P < 0.05). Although the viability, motility and acrosome integrity of sperm cryopreserved with a glucose-containing extender did not significantly differ from sperm frozen in the sorbitol-based extender when examined at 2 and 4 h post-thaw, all of these parameters plus plasma membrane functionality were improved for sperm frozen in the sorbitol extender than in the glucose extender when examined 10 min post-thaw. Two of four mares (50%) inseminated with semen frozen with a sorbitol-containing freezing extender became pregnant. It is concluded that different sugars have different abilities to protect against cryoinjury during freezing and thawing of stallion sperm. This study demonstrated that an extender containing sorbitol as primary sugar can be used to successfully cryopreserve equine sperm; moreover, the quality of frozen-thawed sperm appeared to be better than when glucose or fructose was the principle sugar in the freezing extender.

Postpartum ovarian activity and serum estradiol-17beta level in Thai crossbred native mares.

To study the postpartum ovarian activities for investigation of first postpartum oestrus, twenty-five Thai crossbred native mares were monitored after parturition by oestrous detection, transrectal palpation and reproductive ultrasonography. Blood samplings were also taken for estradiol-17beta (E2) analysis. The first ovulation occurred within 20 days postpartum in 92% (23/25) of the mares. The mean intervals of foaling to first oestrus and to first ovulation were 10.3 +/- 2.9 and 13.4 +/- 2.6 days (mean +/- SD) respectively. Serum E2 increased from 7.0 +/- 2.9 to a peak of 10.8 +/- 3.3 pg/ml (mean +/- SD) at 2 days before ovulation. In conclusion, from the study, it can be stated that the postpartum breeding management should be considered after day 10 postpartum by careful examination of ovarian activity with various methods. However, the uterine condition should be also estimated associated with the ovarian activity after parturition which may increase breeding performance and foal production.

Transcervical catheterization and cervical patency during the oestrous cycle in domestic cats.

The aims of the present study were to develop a device for vaginal and transcervical catheterization in domestic cats, and to study cervical patency during the various stages of the oestrous cycle. Seventeen queens submitted for routine spaying were included in the study. A vaginal catheter was designed from a urinary catheter for dogs, to fit into the ventral vaginal fornix, and a 3.5 French tomcat catheter was used as an inner transcervical catheter. Cervical patency was studied by infusing 0.5 ml Urografin into the cranial vagina and taking X-rays of the queens after 5 min. The Urografin did not enter the uterus, even in the oestrous queens. Transcervical catheterization was then attempted. The correct placement of the intrauterine catheter was confirmed by injecting green food colour mixed with penicillin G and observing the presence of stain in the uterine horns during surgery. Catheterization was successful in 13 of 17 queens: six of nine in interoestrus, three of three in oestrus, one of two in metoestrus and three of three in the postpartum period. Transcervical catheterization is a non-invasive technique that is likely to improve the success rate of assisted feline reproduction, and is potentially a useful non-surgical technique for diagnosis and therapy of uterine diseases.