Tick-borne haemoparasite infections are a major challenge in small ruminant (SR) production across tropical areas. The present study evaluated the prevalence of Theileria, Babesia and Anaplasma in SRs and their tick vectors and estimated the association between pathogen prevalence with clinical hematological findings among SR populations in Kurdistan province, western Iran. In total, 250 blood samples and 250 tick species (one per animal) were collected from SR populations, along with clinical and hematological examinations. Microscopy of blood smears and molecular analysis were performed to detect potential infection with Theileria, Babesia and Anaplasma. Moreover, haemoparasites were explored in the isolated ticks using semi-nested PCR. Based on microscopy, the prevalence of Theileria, Anaplasma and Babesia infections was 91.2%, 23.2% and 2.4%, respectively. Semi-nested PCR analysis of blood samples demonstrated 86.8%, 78.8% and 14% prevalence for T. ovis, A. ovis and B. ovis, respectively. Dermacentor marginatus and Rhipicephalus turanicus were predominant isolated tick vectors from SR, while D. marginatus was the most contaminated tick in all investigated counties. There were, also, a statistically significant association between the estimated molecular prevalence rates with semi-yellow conjunctiva (A. ovis), body temperature (T. ovis and A. ovis), heart rate (T. ovis and B. ovis), mean white blood cell count (T. ovis and A. ovis), mean red blood cell count (T. ovis and B. ovis), as well as mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration in all haemoparasite infections. Future studies are recommended to reveal the epidemiology of such infections in SRs in Iran.

Method validation is a cornerstone on which biomarker development and utilization rest. However, given the abundance of biomarker candidates that are being identified and characterized, validation of these entities for the use in nonclinical studies can be complex. The objective of this continuing education course was to review current practices and challenges encountered during the validation of methods for the analysis of novel biomarkers. Additionally, the importance of biological validation and correlation with pathology end points for biomarker candidates was discussed. This article is a summary of the materials presented at the 36th Annual Symposium of the Society of Toxicologic Pathology for a continuing education course titled "Current Practices and Challenges in Method Validation." The speakers were subject-matter experts in the validation of quantitative mass spectrometry, multiplex binding assays, biological biomarkers, and immunophenotyping and anatomic and clinical pathology considerations in biomarker qualification.

The attempts via introducing many methods have been conducted to select the best antibiotic combination in the treatment of seriously ill patients. Operational or interpretational complexity or time-consuming along with sufficient accuracy led to postpone routine clinical use of these tests until today, despite the urgent need for them. By this study and proposed method, selection of the best double antibiotic synergistic combination against resistant pathogen is simply same as Kirby-Bauer antibiotic susceptibility test. It seems, precise and reliable results (very low coefficient of variation) will be introduced it as a routine accurate diagnostic doubled antimicrobial synergism test.•The objective of this study was to introduce a novel method in antibiotic interaction detection.•It demonstrates high sensitivity and accuracy.•Easy implementation by routine microbiology labs materials and equipment and so easy stand-alone interpretation seems to make it friendly test be able to replacing the previous methods.

Standard components of nonclinical toxicity testing for novel pharmaceuticals include clinical and anatomic pathology, as well as separate evaluation of effects on reproduction and development to inform clinical development and labeling. General study designs in regulatory guidances do not specifically mandate use of pathology or reproductive end points across all study types; thus, inclusion and use of these end points are variable. The Scientific and Regulatory Policy Committee of the Society of Toxicologic Pathology (STP) formed a Working Group to assess the current guidelines and practices on the use of reproductive, anatomic pathology, and clinical pathology end points in general, reproductive, and developmental toxicology studies. The Working Group constructed a survey sent to pathologists and reproductive toxicologists, and responses from participating organizations were collected through the STP for evaluation by the Working Group. The regulatory context, relevant survey results, and collective experience of the Working Group are discussed and provide the basis of each assessment by study type. Overall, the current practice of including specific end points on a case-by-case basis is considered appropriate. Points to consider are summarized for inclusion of reproductive end points in general toxicity studies and for the informed use of pathology end points in reproductive and developmental toxicity studies.

Standard components of nonclinical toxicity testing for novel pharmaceuticals include clinical and anatomic pathology, as well as separate evaluation of effects on reproduction and development to inform clinical development and labeling. General study designs in regulatory guidances do not specifically mandate use of pathology or reproductive end points across all study types; thus, inclusion and use of these end points are variable. The Scientific and Regulatory Policy Committee of the Society of Toxicologic Pathology (STP) formed a Working Group to assess the current guidelines and practices on the use of reproductive, anatomic pathology, and clinical pathology end points in general, reproductive, and developmental toxicology studies. The Working Group constructed a survey sent to pathologists and reproductive toxicologists, and responses from participating organizations were collected through the STP for evaluation by the Working Group. The regulatory context, relevant survey results, and collective experience of the Working Group are discussed and provide the basis of each assessment by study type. Overall, the current practice of including specific end points on a case-by-case basis is considered appropriate. Points to consider are summarized for inclusion of reproductive end points in general toxicity studies and for the informed use of pathology end points in reproductive and developmental toxicity studies.

The purpose of this paper by the Regulatory Affairs Committee (RAC) of the American Society for Veterinary Clinical Pathology (ASVCP) is to review the current regulatory guidances (eg, guidelines) and published recommendations for best practices in veterinary toxicologic clinical pathology, particularly in the pharmaceutical and biotechnology industries, and to utilize the combined experience of ASVCP RAC to provide updated recommendations. Discussion points include (1) instrumentation, validation, and sample collection, (2) routine laboratory variables, (3) cytologic laboratory variables, (4) data interpretation and reporting (including peer review, reference intervals and statistics), and (5) roles and responsibilities of clinical pathologists and laboratory personnel. Revision and improvement of current practices should be in alignment with evolving regulatory guidance documents, new technology, and expanding understanding and utility of clinical pathology. These recommendations provide a contemporary guide for the refinement of veterinary toxicologic clinical pathology best practices.

This study was conducted to determine the prevalence and antimicrobial resistance of Campylobacter spp. isolated from retail raw meats in Iran. From August 2009 to August 2010, a total of 379 raw meat samples from camel (n = 130), beef (n = 207), and water buffalo (n = 42) were purchased from randomly selected retail outlets in Chaharmahal va Bakhtiari and Khuzestan provinces in Iran. The samples were evaluated for the presence of Campylobacter using traditional bacteriological tests and a nested polymerase chain reaction. Overall, 31 of 379 meat samples (8.2%) were contaminated with Campylobacter. The highest prevalence of Campylobacter spp. was found in water buffalo meat (21.4%), followed by beef (9.2%), and camel (2.3%) meat. The most prevalent Campylobacter species isolated from meat samples was Campylobacter jejuni (77.4%); the remaining isolates were Campylobacter coli (22.6%). Susceptibilities of 31 Campylobacter isolates were determined for ten antimicrobial drugs using the disk diffusion assay. Of 31 Campylobacter isolates, 27 (87.1%) were resistant to one or more antimicrobial agents. Nine strains (29.0%) were resistant to one single antimicrobial agent, and eight strains (25.8%) showed resistance to two antimicrobial agents. Multidrug resistance was found in 32.3% of Campylobacter strains. Resistance to tetracycline was the most common finding (67.7%), followed by resistance to ciprofloxacin (32.7%), and nalidixic acid (32.7%). To the authors' knowledge, the present study is the first report of the isolation of Campylobacter spp. from raw water buffalo meat in Iran.

A total of 150 bovine (60), ovine (42), and caprine (48) bulk milk samples were analyzed using a commercially available competitive ELISA kit. Overall, AFM1 was found in 46.7 % of the analyzed samples by an average concentration of 40.3 ± 22.2 ng/L. The incidence rates of AFM1 contamination in bovine, ovine, and caprine bulk milk samples were 66.7, 31.0, and 35.4 %, respectively. The concentration of AFM1 in 37.5 % of AFM1-positive bovine milk samples and 5.9 % of AFM1-positive caprine milk samples were higher than 50 ng/L.

Currently, somatic cell count (SCC) and bacterial culture are considered as the gold standard methods for the diagnosis of bovine subclinical mastitis. However, SCC has a low diagnostic accuracy. Therefore, for identification of infected animals, new biomarkers with high diagnostic accuracy are needed. Acute phase proteins (APPs) are proteins that are increased (positive APPs) or decreased (negative APPs) in response to inflammation. The objective of this study was to determine the diagnostic value of milk APPs for the diagnosis of subclinical mastitis in dairy cows. A total of 90 clinically healthy cows were randomly selected. Of these, 52 cows were considered subclinical mastitic based on a SCC higher than 130 × 1,000 cells/mL of milk and positive bacterial culture results of milk samples obtained from at least one of the quarters. Milk amyloid A (MAA) concentration was measured using a commercial ELISA kit and albumin, α-lactalbumin, β-lactoglobulin, and immunoglobulin (Ig) were measured in the whey samples by the use of cellulose acetate electrophoresis. Diagnostic sensitivity and specificity and cutoff points for each test were determined via receiver-operating characteristics analysis. Significant (P < 0.001) increases in the mean and median concentration of MAA, albumin, α-lactalbumin, and Ig were found in the milk samples collected from cows with subclinical mastitis. MAA was the most accurate test with a diagnostic sensitivity of 92.3% and specificity of 92.1% at cutoff point of >1.6 mg/L. The results of this study showed that determination of MAA can be used as a reliable method for the diagnosis of bovine subclinical mastitis.

A 4-year old, male cynomolgus monkey (Macaca fascicularis) imported from China was acquired by Wyeth Research from a commercial source. On physical examination the animal had a 2 cm in diameter pigmented raised mass at the dorsal aspect of the neck. The mass was surgically removed for cytologic and histopathologic evaluations. Touch imprints of the mass stained with Wright–Giemsa revealed pleomorphic neoplastic cells arranged individually and in small clusters, admixed with extracellular pigment granules. The neoplastic cells were mostly round to oval and were filled with green-black to blue-black pigment granules obscuring the nucleus. The pigment granules were about 1 μm, oval to elongate, and consistent with melanin granules. Histologically, this nonencapsulated mass was composed of poorly demarcated, heavily pigmented melanocytes diffusely infiltrating the dermis. In the bleached sections, the neoplasm was composed of fairly uniform round to polygonal cells arranged in packets and supported by a fibrovascular stroma. The cells had discrete cell borders, and the nuclei were mostly round with finely stippled chromatin and lacked nuclear atypia. The cytologic and histologic findings supported a final diagnosis of melanocytoma. To the authors’ knowledge, the present case is the first cytologic description of a cutaneous melanocytoma in a nonhuman primate.

A 13-year-old, male, North American elk (Cervus elaphus canadensis) from a commercial herd was presented for a sudden onset of lethargy, anorexia, and hematuria to Palmer Veterinary Clinic, Plattsburgh, NY, USA. The elk, originated from Ohio, was from a herd of 60 animals with an age range of 2 months to 13 years. Clinically, over a 5–7-day period, the elk began to distance itself from the remainder of the herd, and became anorectic and eventually recumbent. Results of the admitting complete blood count indicated a normocytic, normochromic, nonregenerative anemia. The animal was serologically negative for elk wasting disease. Supportive treatment did not result in clinical improvement. The elk was euthanized because of the rapidly declining condition. At necropsy, the spleen was diffusely enlarged and the mesenteric and periaortic fats were yellow. Microscopically, the only finding of note was extensive splenic erythrophagocytosis. Microscopic examination of a peripheral blood film stained with Wright–Giemsa revealed that about 5% of erythrocytes contained protozoal parasites. The morphologic characteristics of these organisms were consistent with Babesia organism, most likely Babesia odocoilei. Using a nested polymerase chain reaction, the organism was identified as B. odocoilei. To the authors’ knowledge, this is the first report of B. odocoilei infection in North American elk in New York State, USA. We recommend that infection with B. odocoilei be considered in any case of acute hemolytic anemia in elk.

BACKGROUND:The time from sampling to analysis can be delayed when blood samples are shipped to distant reference laboratories or when analysis cannot be readily performed.OBJECTIVE:The objective of this study was to evaluate the stability of hematologic analytes in blood samples from monkeys, rabbits, rats, and mice when samples were stored for up to 72 hours at 4°C.METHODS:Blood samples from 30 monkeys, 15 rabbits, 20 rats, and 30 mice were collected into EDTA-containing tubes and were initially analyzed within 1 hour of collection using the ADVIA 120 analyzer. The samples were then stored at 4°C and reanalyzed at 24, 48, and 72 hours after collection.RESULTS:Significant (P<.0003) changes in hematologic analytes and calculations included increased HCT and MCV and decreased MCHC and cell hemoglobin concentration mean (CHCM) at 72 hours and increased MPV at 24 hours in monkeys; increased MCV at 72 hours and MPV at 48 hours and decreased monocyte count at 24 hours in rabbits; increased MCV and decreased MCHC, CHCM, and monocyte count at 24 hours in rats; increased MCV, red cell distribution width, and MPV and decreased MCHC, CHCM, and monocyte count at 24 hours in mice.CONCLUSIONS:Although most of the changes in the hematologic analytes in blood from monkeys, rabbits, rats, and mice when samples were stored at 4°C were analytically acceptable and clinically negligible, the best practice in measuring hematologic analytes in these animals is timely processing of blood samples, preferably within 1 hour after collection.

The most widely used method for bilirubin concentration determination is the diazo method, which measures the color of azobilirubin. The vanadate oxidase method is based on oxidation of bilirubin to biliverdin by vanadate. The objective of this study was to compare total and direct bilirubin concentration ([Bt] and [Bd], respectively) determined by the diazo and vanadate oxidase methods in pooled serum samples from dogs, monkeys, and rats spiked with panels of different concentrations of bilirubin standards. Pooled serum samples from 40 dogs, 40 monkeys, and 60 rats were spiked with either ditaurine conjugates of bilirubin or a standard reference material. The results obtained from both assays were compared using Deming regression analysis. The intra- and interassay precision, expressed as a percentage of the coefficient of variation (%CV), was determined for [Bt] and [Bd], and the mean percentage of recovery was calculated. The vanadate oxidase method displayed an excellent correlation (r = 0.99–1.00) with the diazo method. Using Deming regression, there were minimal negative or positive constant and proportional biases for [Bt] and [Bd]. The precision studies revealed that the vanadate oxidase method has comparable between-run and within-run CVs to those of the diazo method. The recovery study demonstrated that the diazo method more closely approximates the expected values of [Bt]. In conclusion, the vanadate oxidase method is a simple and rapid method that can be employed as an alternative to the diazo method when interfering substances are present in the serum samples of dog, monkey, and rat.

An 8-year-old stallion male Caspian miniature horse was acquired by the Veterinary Teaching Hospital, Islamic Azad University from a commercial source for teaching veterinary students. One year later, during an annual health screen, a mass was noted in the perineal region. The mass was surgically removed. Histologic examination revealed that the mass was a relatively well-demarcated, unencapsulated neoplasm composed of streams, bundles, and whorls of neoplastic spindle cells that had oval to fusiform nuclei with stippled chromatin, variably distinct nucleolus, and moderate amounts of eosinophilic cytoplasm. Based on the morphologic and histologic findings, a diagnosis of equine sarcoid was made. Results of bacteriologic tests and fungal culture of the mass were negative. By re-examining the horse 3 weeks after surgery, the affected site was completely healed. To the authors’ knowledge, this is the first report of sarcoid in a Caspian miniature horse.

This study was conducted to determine the prevalence and antimicrobial resistance pattern of Campylobacter spp. isolated from retail raw poultry meats in Iran. From July 2009 to March 2010, a total of 494 raw meat samples from chicken (n = 200), turkey (n = 170), quail (n = 86), partridge (n = 17), and ostrich (n = 21) were purchased from randomly selected retail outlets in Shahrekord, Iran. Using cultural method, 187 meat samples (37.9%) were contaminated with Campylobacter. The highest prevalence of Campylobacter spp. was found in chicken meat (47.0%) followed by quail (43.0%), partridge (35.3%), turkey (28.8%), and ostrich (4.8%) meat. The most prevalent Campylobacter species was Campylobacter jejuni (92.0%). The PCR assay could identify 38 Campylobacter-contaminated samples that were negative using the cultural method. Antimicrobial susceptibility test results showed that 98.4% of isolates were resistant to one or more antimicrobial agents. Resistance to tetracycline was the most common findings (70.6%), followed by resistance to nalidixic acid (54.0%), and ciprofloxacin (49.7%). Significantly higher prevalence rates of Campylobacter spp. (P < 0.05) were found in meat samples taken in summer (51.1%). To our knowledge, the present study is the first report of the isolation of Campylobacter spp. from raw partridge meat in Iran. (C) 2011 Elsevier Ltd. All rights reserved.