Comparison of Whole Ovary Cryotreatments for Fertility Preservation

The goal of this study was to compare a traditional slow‐freeze method (TF) with an open unidirectional slow freeze cooling system (UF) for whole ovary cryopreservation. Therefore, whole pig ovaries were randomly assigned to (A) fresh control, (B) traditional slow freeze (TF) or (C) unidirectional slow freeze (UF). Ovaries were perfused with 10% DMSO in Krebs‐Ringer. For TF, whole ovaries were placed in specimen jars containing 10% DMSO and placed into a specialized container for freezing filled with propan‐2‐ol. For UF, whole ovaries were placed within a specially designed container containing 10% DMSO and transferred to a specialized freezing machine (CTE 920). Histological evaluation demonstrated intact morphology of follicles in all groups; however, an overall decrease of follicle numbers in TF (46%) and UF (50%) compared to fresh control. Live/dead assay indicated significantly lower populations of live cells in both TF (60%) and UF (58%) compared to fresh tissue (74%). TUNEL assay confirmed a difference in percentage of apoptotic follicles between fresh and TF, but there was no significant difference between fresh and UF. To improve the structural and functional integrity of whole ovaries, further investigation, especially into directional freezing, is needed. Whole ovary cryopreservation could provide opportunities for women facing fertility loss due to chemo‐ or radiotherapy treatment.     

Gastrointestinal pythiosis with concurrent presumptive gastrointestinal basidiobolomycosis in a Boxer dog

A 2‐year‐old female spayed Boxer dog was presented for a 1‐month history of progressive hemorrhagic diarrhea with tenesmus and weight loss despite trial courses of antibiotics and diet change. Abdominal ultrasound revealed severe, focal thickening, and loss of normal architecture of the colonic wall with abdominal lymphadenomegaly. Dry‐mount fecal cytology, performed on several consecutive days, consistently revealed numerous, round, 16‐20 μm structures with basophilic, granular content, and a thin cell wall. Transmission electron microscopy identified these structures as fungi. Culture, polymerase chain reaction (PCR), and sequencing of the internal transcribed spacer, D1/D2 regions, and DNA‐directed RNA polymerase II core subunit (RPB2) confirmed the presence of Basidiobolus microsporus in the feces. Biopsies collected via ileocolonoscopy revealed marked, multifocal, chronic, neutrophilic, and eosinophilic ileitis and colitis with ulceration, granulation tissue, and intralesional hyphae (identified with Gomori methenamine silver stain). A Pythium enzyme‐linked immunosorbent assay and Pythium‐specific PCR performed on the formalin‐fixed paraffin‐embedded biopsy specimens were positive while Basidiobolus‐specific PCR was negative, thus confirming a diagnosis of pythiosis. This report describes a fatal case of colonic and intestinal pythiosis with the presence of fecal Basidiobolus sp. spores, suggestive of concurrent gastrointestinal basidiobolomycosis.     

Natural Heterobilharzia americana infection in horses in Texas

The schistosome Heterobilharzia americana infects dogs, raccoons, and other mammals in the southeastern United States. Migration of eggs into the liver results in parasitic granulomas with varying degrees of fibrosis and inflammation. Recently, hepatic parasitic granulomas in horses were shown to be caused by H. americana infection. In the present study, samples of liver from 11 of 12 horses with hepatic granulomas identified at necropsy (n = 11) or surgical biopsy (n = 1) were used for DNA extraction, polymerase chain reaction amplification and sequencing using primers specific for a portion of the H. americana small subunit ribosomal RNA gene. A polymerase chain reaction amplicon of the correct size was produced from the extracted DNA in 8 of the 11 horses. Amplicons from 5 of the 8 positive horses were sequenced and had 100% identity with H. americana. In all but 2 of the 12 horses, Heterobilharzia was not responsible for the primary clinical disease, and the hepatic granulomas were considered an incidental finding.     

A comparative study of the effect of 2 hormonal treatment protocols on the reproductive performance of previously anoestrous dairy cows

The objective of this study was to compare the reproductive performance of anoestrous dairy cows treated just prior to the mating start date (MSD) with a Controlled Internal Drug Releasing Device (CIDR) which was placed intravaginaly for either 6 or 8 days, and a combination of oestradiol benzoate. Lactating dairy cows (n = 926) that had been diagnosed with anovulatory anoestrus were divided into two sub-groups. A hormonal treatment protocol that involved the use of a CIDR device containing 1.9 g of progesterone was inserted into the vagina of each cow and left intravaginally for either 6 (6-day group, n = 441) or 8 days (8-day group, n = 485). Every cow in the trial was inseminated after being detected in oestrus from Day -2 onwards (where Day 0, was a herd's MSD), using an appropriate detection aid according to the herds' preference. Cows that had been seen in oestrus and were inseminated by Day 2 were selected for re-synchrony. The standardised re-synchrony involved the re-insertion of a previously used CIDR device into the vagina of each cow on Day 14, together with an injection of 1.0 mg oestradiolbenzoate i.m. This CIDR device was removed on Day 22 and each of these cows injected with 1.0 mg oestradiol benzoate i.m. on Day 23. Each re-synchronised cow that was detected in oestrus was re-inseminated. Treatment with an 8-day CIDR increased the proportion of cows submitted for insemination within the first 3 days of the MSD, compared with the 6-day group (83.7 % vs 71.2%, respectively, P < 0.001), as well as the proportion of cows conceiving within the first 3 days of MSD (36.2% vs 27.7%, P = 0.02), but reduced both the interval from MSD to the first service (4.5 +/- 0.5 vs 6.8 +/- 0.7, P = 0.01), and the interval from MSD to conception (28.1 +/- 1.5 vs 34.0 +/- 1.8, P = 0.009). A greater percentage of the cows in the 6-day group that were not pregnant to the first insemination were submitted for a second insemination by Day 28 compared with the 8-day group (81.1% vs 68.3%, P < 0.001). Conception rates for cows submitted for this second insemination by Day 28 of the MSD were also higher in the 6-day than in the 8-day group (48.4% vs 33.9%, P = 0.009). The percentage of cows pregnant at the end of a herd's AI period of 6 weeks did not differ (57.1% vs 54.8% for 8-day and 6-day groups, respectively, P = 0.42); neither did the proportion of cows pregnant at the end of the a herd's combined AI and natural mating period of 21 weeks (81.4% vs 79.2%, for 8-day and 6-day groups respectively P = 0.36). Treatment of anovulatory anoestrous dairy cows with a combination of an 8-day CIDR and oestradiol benzoate before the MSD improved their reproductive performance by increasing the portion of cows submitted for insemination within the first 3-days of the MSD by reducing the interval from MSD to first service and by increasing the conception rate to the first insemination to collectively reduce the average interval from MSD to conception.     

The spread of Corynebacterium pseudotuberculosis infection to unvaccinated and vaccinated sheep

SUMMARY The decrease in the prevalence of Corynebacterium pseudotuberculosis after two generations of vaccination against the disease it causes, was used to estimate the rate of control of caseous lymphadenitis (CLA). Three groups of 150 sheep, of which 50 in each group were artificially infected with C pseudotuberculosis and 100 in each group were uninfected sheep, were run separately for 40 months and shorn 5 times to promote the spread of CLA. One lot of 50 infected sheep and 2 lots of 100 uninfected sheep were vaccinated against CLA. The rate of spread of CLA was recorded. Sheep vaccinated against CLA and naturally exposed to infection had a 74% lower infection rate than unvaccinated sheep. Sheep vaccinated against CLA and exposed to only vaccinated infected sheep had a 97% lower infection rate. Unvaccinated sheep had a 76% infection rate, with 77% of the transmission occurring at the 4th and 5th shearings, without any discharging CLA abscesses being observed. This study supports the view that in Australian wool producing flocks, CLA spreads mainly from sheep with discharging lung abscesses to sheep with shearing cuts. Vaccinated sheep infected with CLA have 96% fewer lung abscesses compared with unvaccinated infected sheep and are therefore less likely to spread this disease to other sheep .     

Effect of Tissue Processing on Assessment of Endoscopic Intestinal Biopsies in Dogs and Cats

Background: Prior studies failed to detect significant association between hypoalbuminemia and small intestinal lesions.
Hypothesis: Use of pictorial templates will enhance consistency of interpathologist interpretation and identification of intestinal lesions associated with hypoalbuminemia.
Animals: Tissues from 62 dogs and 25 cats examined as clinical cases at 7 referral veterinary practices in 4 countries.
Methods: Retrospective, observational study. Histopathology slides from sequential cases undergoing endoscopic biopsy were examined by 4 pathologists by pictorial templates. Changes for 9 microscopic features were recorded as normal, mild, moderate or severe, and 2- and 4-point scales were tested for consistency of interpretation. Logistic regression models determined odds ratios (OR) of histologic lesions being associated with hypoalbuminemia while κ statistics determined agreement between pathologists on histologic lesions.
Results: There was poor agreement (κ=−0.013 to 0.3) between pathologists, and institution of origin of slides had effect (κ= 1.0 for 3 of 4 lesions on slides from Institution 5) on agreement between pathologists on selected histologic features. Using 2 point as opposed to 4-point grading scale increased agreement between pathologists (maximum κ= 0.69 using 4-point scale versus maximum κ= 1.0 using 2-point scale). Significant association ( P = .019– .04; 95% OR = 3.14–10.84) between lacteal dilation and hypoalbuminemia was found by 3 pathologists.
Conclusions and Clinical Importance: Substantial inconsistency between pathologists remains despite use of pictorial template because of differences in slide processing. Distinguishing between mild and moderate lesions might be important source of the disagreement among pathologists.