Use of a Point-of-Care Progesterone Assay to Predict Onset of Parturition in the Bitch.

An assay of circulating progesterone (P4) is commonly used to estimate progress through late gestation in the bitch. Point-of-care assays provide rapid results, a major advantage over laboratory-based assays. This study aims to compare P4 levels determined by the Catalyst® Progesterone point-of-care assay with those determined by chemiluminescent immunoassay (CLIA) and to identify the expected distribution of Catalyst P4 levels at time intervals 3 days prior to the onset of parturition in pregnant bitches. Twenty-eight pregnant bitches carrying two or more fetuses were admitted to a specialist veterinary reproduction hospital 53 days after the onset of cytological diestrus or, when that date was not known, 57 days after the last mating. Vaginal speculum examinations were performed every 6 h until the onset of cervical dilatation (TCD). Serum samples were collected twice daily (08h00 and 18h00) until TCD. For most samples, fresh serum was assayed for P4 immediately using the Catalyst assay (CatP4), then frozen until assayed by CLIA (IMMULITE 2000; ImmP4). However, for some samples, CatP4 was not analyzed prior to freezing. For these data points (n = 33), CatP4 for fresh serum was estimated from CatP4 assayed on frozen-thawed serum, based on a comparison between CatP4 on fresh vs. frozen-thawed sera. In comparison to ImmP4, CatP4 levels up to and including 7 nmol/L appear to have a constant bias of -1.69 nmol/L (limits of agreement -4.91 to 1.52), while levels >7 nmol/L appear to have a proportional bias of -17.9% (limits of agreement -68.6% to 32.7%). Bootstrapped percentiles of CatP4 results spanned 0.4-9 nmol/L within 12 h of TCD, 0.9-11 nmol/L 12-24 h from TCD, and 2.2-13.5 nmol/L 24-36 h from TCD. A CatP4 >9 nmol/L indicates a bitch that is unlikely to reach TCD within 12 h. Bitches with CatP4s below 3.5 nmol/L are likely to reach TCD within 36 h and bitches with a CatP4 below 2.2 nmol/L are likely to reach TCD within 24 h. In conclusion, the Catalyst Progesterone assay provides rapid assessment of circulating P4 in the bitch, with clinical application in the monitoring of late term pregnant bitches.

Effects of ivermectin treatment of backyard chickens on mosquito dynamics and West Nile virus transmission.

BACKGROUND: Vector control strategies typically rely on pesticides to target mosquitoes involved in enzootic and zoonotic transmission of West Nile virus (WNV). Nevertheless, increasing insecticide resistance and a desire to reduce pesticide usage provide the impetus for developing alternative strategies. Ivermectin (IVM), an antiparasitic drug which is widely used in human and veterinary medicine, is a potential alternative for targeted control because Culex mosquitoes experience increased mortality following ingestion of IVM in bloodmeals. METHODOLOGY/PRINCIPAL FINDINGS: We conducted a randomized field trial to investigate the impact of treating backyard chicken flocks with IVM in urban neighborhoods across Davis, California on mosquito populations and WNV transmission dynamics. We observed a significant reduction in WNV seroconversions in treated vs. untreated chickens, suggesting a reduction in WNV transmission intensity around treated flocks. We also detected a reduction in parity rates of Cx. tarsalis near treated vs. untreated flocks and increased mortality in wild mosquitoes following a bloodmeal on treated chickens (IVM serum concentration > 5ng/mL) vs. chickens with IVM serum concentrations < 5 ng/mL. However, we did not find a significant difference in abundance or infection prevalence in mosquitoes between treatment groups associated with the reductions in seroconversions. Mosquito immigration from surrounding larval habitat, relatively low WNV activity in the study area, and variable IVM serum concentrations likely contributed to uncertainty about the impact. CONCLUSIONS/SIGNIFICANCE: Taken together, our results point to a reduction in WNV transmission due to the impact of IVM on Culex mosquito populations and support the ongoing investigation of oral administration of IVM to wild birds for local control of WNV transmission, although further work is needed to optimize dosing and understand effects on entomological endpoints.

Use of serum progesterone and prostaglandin F2α metabolite levels to predict onset of parturition in the bitch.

The prediction of time to onset of parturition in a preparturient bitch is of great clinical value, particularly for bitches at high risk of dystocia and those lacking relevant clinical data from the time of breeding. In a previous study, four cut-offs for plasma progesterone levels, measured by radioimmunoassay, were shown to be useful for predicting the likelihood of a bitch entering stage one of parturition within defined time intervals. The first aim of the current study was to evaluate these cut-offs in a clinical setting, using serum progesterone samples drawn from preparturient bitches 12-hourly instead of 6-hourly and assessed using chemiluminescence immunoassay. Furthermore, the use of 13,14-dihydro-15-keto-prostaglandin F2α, (PGFM), a metabolite of prostaglandin F2α , in predicting the time to onset of parturition was evaluated. Forty bitches carrying two or more foetuses were admitted to a specialist veterinary reproduction hospital 53 d after the onset of cytological dioestrus when that date was known, or 57 d after the last mating. Vaginal speculum examinations were performed every 6 hr until cervical dilatation was visualized (time of cervical dilatation; TCD). Serum samples were collected at 08h00 and 18h00 daily until TCD. All bitches underwent elective caesarean section at TCD. Results of this study show that approximately 5% and 10% of preparturient bitches will reach TCD within 12 hr despite a serum progesterone level of at least 15.8 nmol/L and 8.7 nmol/L respectively. In addition, there is a 95% probability that a preparturient bitch will reach TCD within 48 hr if her serum progesterone level is below 8.7 nmol/L, and a 91% probability of her reaching TCD within 24 hr if her serum progesterone level is below 3.18 nmol/L. Approximately 90% of bitches that demonstrate a 20% increase in PGFM over a 12-hr period are likely to be within 36 hr of TCD. These results provide useful benchmarks for the management of canine parturition.

Towards scheduled pre-parturient caesarean sections in bitches.

Elective caesarean section (CS) is the safest means of delivering the litter in bitches in specific situations. Timeously performing elective pre-parturient CSs at a fixed time would be convenient and prevent emergency CSs and foetal demise. This review proposes a method of performing elective pre-parturient CSs which is safe for both the bitch and puppies. Brachycephaly, small litters and large litters, preceding litter delivered by CS and trial of labour after a preceding CS are identified as factors increasing the need for CS whereas emergency CS is identified as factor increasing foetal demise. The first day of cytological dioestrus more precisely predicts the day of onset of spontaneous parturition than the first day of the LH surge or the dates during oestrus on which progesterone (P4) first exceeds 6 nM or 16 nM. Foetal biparietal diameter at the time of onset of spontaneous parturition varies too much to accurately predict readiness for CS. During the last few days of gestation, P4 with cut-off concentrations at 15.8, 8.7 and 3.18 nM, but not plasma cortisol concentrations, hold promise as predictors of onset of parturition and when to perform pre-parturient CSs. A protocol associating medetomidine hydrochloride as premedicant with propofol as induction agent and sevoflurane as maintenance is safe for scheduled CS and yields good maternal and puppy survival rates at delivery, 2 hr and 7 days after CSs. Clinicians have to pay attention to the haematocrit of bitches at the time of cervical dilatation which is at the lower end of the normal reference ranges for non-pregnant dogs and to the decline in haematocrit during CS (as a proxy for blood loss) which is approximately 7% for both parturient (open cervix) and pre-parturient (closed cervix) CSs. Pre-parturient CSs can be scheduled and performed 57 days after onset of cytological dioestrus with puppy survival rates of 99%. Collectively, these studies provide a protocol to safely perform elective CSs in a large proportion of the obstetric population at a convenient time of the day but more research is required with larger numbers to establish whether this practice is routinely safe and safe in all breeds.

Comparison of progesterone assay by chemiluminescence or radioimmunoassay for clinical decision-making in canine reproduction.

The Coat-A-Count® radioimmunoassay has been long and widely used to determine the concentration of progesterone in serum or plasma of bitches (progRIA), but was discontinued in 2014. The Immulite® 1000 LKPG1 chemiluminescence immunoassay has gained prominence since 2003 to determine the concentration of progesterone in serum of bitches, but the assay changed in 2012 (Immulite® 1000 LKPW1). This study assessed the feasibility of using Immulite® 1000 LKPW1 (progImm) to estimate the time of clinically relevant events during oestrus and compared progRIA and progImm 2 and 3 days after the first or only day of the luteinizing hormone surge (LH1). ProgImm first exceeded 5.1 nmol/L on the same day that progRIA first exceeded 6 nmol/L, a proxy for the occurrence of the LH surge, or the day before in 28 of 31 (90%) of oestrous periods. ProgImm first exceeded 13.6 nmol/L on the same day that progRIA first exceeded 16 nmol/L (a proxy for the day of ovulation) or the day before in 34 of 35 (97%) oestrous periods. ProgImm first exceeded 5.4 nmol/L on LH1 or the day before in 24 of 25 (95%) of oestrous periods. The median of progImm 2 days after LH1 was 1.2 nmol/L lower than the 10.7 nmol/L of progRIA (p = 0.001). The mean of progImm 3 days after LH1 was 2.2 nmol/L lower than the 19.0 nmol/L of progRIA (p 0.001). In conclusion, the days on which progImm first exceeded 5.1 nmol/L, 13.6 nmol/L and 5.4 nmol/L effectively estimate the days on which progRIA reached 6 nmol/L or 16 nmol/L or LH1.

Comparing the values of progesterone in the blood of bitches as measured with a chemiluminescence immunoassay and a radioimmunoassay.

A 125 I radioimmunoassay (RIA) has long been used to determine the value of progesterone in serum or plasma of bitches but was discontinued in 2014. A chemiluminescence immunoassay (CLIA) gained prominence since 2003 to determine the value of progesterone in serum of bitches but the assay changed in 2012. This study assessed the agreement between progesterone values obtained with RIA in plasma (progRIA) and with the post-2012 CLIA (progCLIA) in the serum of bitches. ProgCLIA was determined in 110 serum samples from 40 bitches in pro-oestrus or early oestrus and compared to progRIA in plasma samples collected from the same bitches at the same time, where progRIA had a uniform distribution between 0.5 and 25 nmol/L. Two replicate analyses of each serum or plasma sample were simultaneously done in the same assay. For RIA and CLIA, the intra-assay CVs were 5.85% and 6.70% and the interassay CVs 8.45% and 9.16%. For RIA and CLIA the progesterone values obtained with replicate analyses differed by as much as 11%-31% in 25% of samples. On average, the value of progCLIA was 85% of that of progRIA (95% CI 58%-112%, n = 110), with 88% of progCLIAs being lower than the progRIAs. This study shows that RIA and CLIA may yield replicate values that differ by as much as 11%-30% in about a quarter of samples analysed, necessitating replicate analyses if precise values are required. The study provides an equation by which to estimate progCLIA from progRIA.

Dizygotic monochorionic canine fetuses with blood chimaerism and suspected freemartinism.

Two full-term canine fetuses were found to share a placenta during Caesarean section. The fetuses were of discordant gender, with apparently normal male and female external genitalia. Genetic analysis of whole-blood samples obtained from each fetus revealed identical DNA profiles, with more than two alleles detected at six loci. Subsequent genetic analysis of myocardial tissue samples revealed dissimilar DNA profiles, with at most two alleles detected per locus. Superimposition of the tissue-derived profiles matched that derived from the blood samples exactly, except for two loci failing to amplify, and hence demonstrated blood chimaerism. Dissection of the abdomen of the male fetus revealed delayed descent of the testes towards the inguinal canals. Macroscopically, the gonads, uterus and vagina were not identifiable on dissection of the female fetus, although vestigial ovarian tissue and a vagina were detected microscopically. The hypoplastic internal reproductive tract of the female fetus was suggestive of freemartinism and is believed to be the first report of this condition in the canine.

Morbidity and mortality following envenomation by the common night adder (Causus rhombeatus) in three dogs.

In South Africa dogs are frequently presented to veterinarians following snakebite. The offending snakes are usually puff adders (Bitis arietans), cobras (Naja spp.) and mambas (Dendroaspis spp.). Night adder (Causus rhombeatus) bites in dogs have not yet been reported in South Africa. This article deals with three cases of dogs bitten by night adders in which extensive tissue damage was noted and one fatality occurred. Night adder bites may be indistinguishable from puff adder bites. Non-specific treatment included addressing the hypovolaemia and swelling. Specific treatment involving immunotherapy using the South African polyvalent antivenom would be ineffective as it does not contain immunoglobulins against night adder venom. Veterinarians should also include night adders as the possible cause of dogs suffering from severe and painful swellings suspected to be due to snakebites.