Investigation of the Role of Androstenedione-19-oic Acid in the Presence of 19-Norandrostenedione in Intact Male Horse Plasma Using Liquid Chromatography–Tandem Mass Spectrometry

A liquid chromatography–tandem mass spectrometry method was developed to confirm the presence of androstenedione-19-oic acid in intact male equine plasma and to show the source of 19-norandrostenedione in equine plasma. Androstenedione-19-oic acid was recovered from acidified plasma by liquid–liquid extraction using methyl tert-butyl ether and separated on an Ace 5 C₈ column. A triple quadrupole mass spectrometer was used to detect the analytes in negative electrospray ionization mode. Limits of detection, quantification, and confirmation of the method were 0.1, 0.5, and 1.0 ng/mL, respectively. The linear dynamic range of quantification was 0.5–50 ng/mL. The presence of androstenedione-19-oic acid was confirmed in all plasma samples obtained from intact male horses but not those from gelded and female horses; the average concentration was 3.1 ± 1.6 ng/mL, suggesting androstenedione-19-oic acid is an endogenous compound only in intact male horse plasma samples. The conversion of androstenedione-19-oic acid to 19-norandrostenedione in equine plasma was demonstrated by spiking androstenedione-19-oic acid into blank plasma and monitoring the generation of 19-norandrostenedione and its increase in concentration during storage. Results indicated that androstenedione-19-oic acid was readily converted into 19-norandrostenedione; the higher the storage temperature, the faster the conversion. The conversion was not affected by the types of plasma samples collected from gelded and female horses or by anticoagulants used in blood collection to harvest plasma. Compared with other matrices such as water, methanol, and phosphate-buffered saline, the conversion of androstenedione-19-oic to 19-norandrostenedione in equine plasma was faster, suggesting that there is an unknown factor(s) in equine plasma that enhances the conversion.     

Effect of post-insemination progesterone supplementation on pregnancy rate in dairy cows

Progesterone plays an important role in maintenance of pregnancy. It is hypothesized that insufficient progesterone early in pregnancy may result in embryonic loss, and that supplemental progesterone would decrease pregnancy loss in dairy cows. In Experiment 1, 84 cows and 16 heifers from a single dairy operation were selected randomly. Within each age category, controlled internal drug release (CIDR) devices were inserted into the vagina of every other female on Day 4 post-insemination and removed on Day 18 post-insemination. Transrectal ultrasonography was performed to determine pregnancy at 4 time periods [days 30 to 37 (week 5), days 44 to 51 (week 7), days 58 to 65 (week 9), and days 86 to 93 (week 13)]. Progesterone supplementation had no effect on pregnancy rate. In Experiment 2, there were no differences in progesterone concentrations between cows that did and did not receive a CIDR. Further, cows receiving CIDR devices did not have an increase in circulating progesterone concentrations 30 min or 1 h after CIDR insertion. It appears that progesterone supplementation does not increase circulating levels of progesterone in the early pregnant lactating dairy cow. Alterative methods to influence progesterone concentrations and/or early embryonic loss need to be investigated.     

Effect of spineless cactus intake (<Emphasis Type="Italic">Opuntia ficus-indica</Emphasis>) on blood glucose levels in lactating sows and its impact on feed intake,body weight loss,and weaning-estrus interval

The effect of spineless cactus intake (Opuntia ficus-indica) on blood glucose (BG) levels in lactating sows and its impact on daily and total feed intake (dFI^?1 and TFI, respectively), body weight loss (BWL), and weaning-estrus interval length (WEI) were evaluated. Thirty-four hybrid (Yorkshire × Landrace × Pietrain) sows in lactation phase were used. Sows were divided into two groups: G1 (n = 17) where they received commercial feed and G2 (n = 17) provided with commercial feed plus an average of 2.0 ± 0.5 kg spineless cactus, based on a sow’s body weight. The variables evaluated were BG, dFI^?1, TFI, BWL, and WEI. Statistical analysis was performed by using a fixed and mixed model methodology, under a repeated measurements experiment. Group effects were found on all analyzed variables (P < 0.05). The BG was lower in G2 (55.2 and 64.5 mg/dL pre- and post-prandial, respectively), compared to that in G1 (70.9 and 80.1 mg/dL pre- and post-prandial, respectively) (P < 0.05). G2 showed better performance than G1 for dFI^?1, BWL, and WEI (P < 0.05) whose averages were 5.5 ± 1.8 kg, 7.4 ± 4.5%, and 5.3 ± 1.2 days, respectively. Averages for these variables in G1 were 4.7 ± 1.5 kg, 16.8 ± 4.6%, and 6.1 ± 1.6 days, respectively. Intake of spineless cactus reduced BG levels in lactating sows, generating greater dFI^?1, lower BWL at the end of lactation, and a lower WEI.     

Fasciola hepatica: Histology of the testis in egg-producing adults of several laboratory-maintained isolates of flukes grown to maturity in cattle and sheep and in flukes from naturally infected hosts

A total of 8 calves approximately 6 months old and 22 lambs of similar age were infected with metacercariae of Fasciola hepatica of various laboratory-maintained isolates including: Cullompton (sensitive to triclabendazole) and Sligo, Oberon and Leon (reported as resistant to triclabendazole). Ten to 16 weeks after infection, flukes were harvested from these experimental animals and the histology of the testis tissue was examined in a representative sample of flukes from each population. Adult wild-type flukes were also collected from 5 chronically infected cattle and 7 chronically infected sheep identified at post-mortem inspection. The testis tissue of these flukes was compared with that of the various laboratory-maintained isolates. Whilst the testes of the wild-type, Oberon and Leon flukes displayed all the usual cell types associated with spermatogenesis in Fasciola hepatica (spermatogonia, spermatocytes, spermatids and mature sperm), the Cullompton flukes from both cattle and sheep showed arrested spermatogenesis, with no stages later than primary spermatocytes represented in the testis profiles. The presence of numerous eosinophilic apoptotic bodies and nuclear fragments suggested that meiotic division was anomalous and incomplete. In contrast to the wild-type flukes, no mature spermatozoa were present in the testes or amongst the shelled eggs in the uterus. A high proportion of the eggs collected from these flukes hatched to release normal-appearing miracidia after an appropriate incubation period, as indeed was the case with all isolates examined and the wild-type flukes. It is concluded that the eggs of Cullompton flukes are capable of development without fertilization, i.e. are parthenogenetic. The implications of this for rapid evolution of resistant clones following an anthelmintic selection event are discussed. Amongst the Sligo flukes examined, two subtypes were recognised, namely, those flukes with all stages of spermatogenesis and mature spermatozoa present in the testes (type 1), and those flukes with all stages of spermatogenesis up to spermatids present, but no maturing spermatozoa in the testes (type 2). Each sheep infected with the Sligo isolate had both type 1 (approximately 60%) and type 2 (approximately 40%) flukes present in the population. Spermatozoa were found amongst the eggs in the uterus in 64% of flukes and this did not necessarily reflect the occurrence of spermatozoa in the testis profiles of particular flukes, suggesting that cross-fertilization had occurred. The apparent disruption of meiosis in the spermatocytes of the Cullompton flukes is consistent with reports that Cullompton flukes are triploid (3n=30), whereas the Sligo and wild-type flukes are diploid (2n=20). In the Sligo flukes the populations are apparently genetically heterogenous, with a proportion of the flukes unable to produce fully formed spermatozoa perhaps because of a failure in spermiogenesis involving elongation of the nucleus during morphogenesis.     

A SURVEY OF CLINICAL MASTITIS IN SOUTH-EAST QUEENSLAND DAIRY HERDS

SUMMARY A survey of clinical mastitis in 26 South-East Queensland dairy herds supplying more than 300,000 litres of milk annually, revealed a quarter incidence of 2.6% with a cow incidence of 9% over a 3-month period. Secretion samples from cows yielded Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, coliforms, Pseudomonas spp and other bacteria in 38.5%, 12.5%, 6.6%, 3.4%, 2.6%. 0.6% and 2.0% of cases respectively. Samples from 38% of the cases were culturally negative while at least 38% of the clinical quarters had shown a previous clinical episode of mastitis. There was a significant association (P < 0.025) between herd and the likelihood of a negative culture result on samples from clinical cases. Twenty-one percent of cases occurred in the first month of lactation and 10.3% in the first week. There was a significant association (P < 0.005) between the age of cow and the stage of lactation in which clinical cases occurred in that cows in the 2 to 5 year age group were more likely to suffer attacks in the earlier part of lactation. There was a highly significant relationship (P < 0.005) between cow age group and the proportion of affected quarters and it was calculated that the change with each year of age was a 55% increase over the previous year. Forty-five percent of the isolates of S. aureus were resistant to penicillin.     

Improvement of trivalent leptospira vaccine by removal of anaphylactic agents

The anaphylactic reactions in cattle following leptospira vaccination mostly booster dose in different parts of Iran have been reported. The serum proteins as allergic substances are components of liquid phase of the vaccine. Therefore, the vaccine was modified by washing the whole cultures by centrifugations. The modified vaccine was safe in laboratory animals and cattle as well as under field conditions. Microagglutination test revealed a similar pattern of antibody response to the three Leptospira interrogans serovars (Canicola, Grippotyphosa, and Sejro hardjo) in all vaccinated cattle groups while was higher than the response of control animals. The results of the present investigation revealed that we can minimize postvaccination shock in vaccinated cattle populations with removing the shock proteins.     

Demographic dynamics and off-take of cattle herds in southern Mali

The herds of 95 families were monitored for 1 year in eight villages in the cotton-growing region of southern Mali. In 2006–2007, reproduction performances were average, with 0.54 calvings/year per cow, and mortality was low. Herd numerical productivity is not very high, less than 0.13/year, because of the high proportion of males kept for animal draught. Depending on the herd size, the behaviour of the families differs, in terms of off-take and in-take of animals. Families that only have one or two draught animals seek to increase their animal draught capacity, with a negative net off-take (−0.13/year). Families with two to three cows have a very low net off-take (0.02/year), with culling of adult animals compensated by purchase. They therefore capitalised this year, with an annual herd growth of 8%. Families with a very large herd (20 to 50 cows) take off more of their stock, with a net off-take of 0.08/year (very few animal purchases) and make a stock growth of 5%. And finally, families with an average-sized herd (6 to 19 cows) take off the whole of the year’s production, with a net off-take of 0.11/year and a nil stock growth rate. The use of a demographic model made it possible to measure the sensitivity of the productivity rate to the different demographic parameters.