Population diversity of Staphylococcus aureus isolated from bovine mastitis in Brazilian dairy herds

Population diversity was evaluated in strains of Staphylococcus aureus isolated from bovine mastitis in Brazilian dairy herds by PCR-RFLP and sequencing of the 3'-terminal portion of the coagulase gene, and the susceptibility of strains to antimicrobials. The results showed great diversity in S. aureus population studied and the existence of predominant clones that account for most infections. No associations between the predominant types observed in the PCR-RFLP and the forms of presentation of the mastitis or to any of the different patterns of antimicrobial resistance were observed.     

Experimental infection with low and high pathogenicity H7N3 Chilean avian influenza viruses in Chiloe wigeon (Anas sibilatrix) and cinnamon teal (Anas cyanoptera)

Two different wild duck species common in Chile and neighboring countries, Chiloe wigeon (Anas sibilatrix) and cinnamon teal (Anas cyanoptera), were intranasally inoculated with 10(6) mean embryo infective dose (EID50) of the H7N3 low pathogenicity (LP) avian influenza virus (AIV) (A/chicken/Chile/176822/02) or high pathogenicity (HP) AIV (A/chicken/Chile/ 184240-1/02), in order to study the infectivity and pathobiology of these viruses. None of the virus-inoculated ducks had clinical signs or died, but most seroconverted by 14 days postinoculation (DPI), indicating a productive virus infection. Both LPAIV and HPAIV were isolated from oral swabs from two of six Chiloe wigeons and from oral and/or cloacal swabs from all five of the cinnamon teal at 2 DPI. Both LPAIV and HPAIV were efficiently transmitted to cinnamon teal contacts but not to Chiloe wigeon contacts. This study demonstrates that the cinnamon teal and Chiloe wigeons were susceptible to infection with both Chilean H7N3 LPAIV and HPAIV, but only the cinnamon teal showed contact transmission of the virus between birds, suggesting that the cinnamon teal has the potential to be a reservoir for these viruses, especially the LPAIV, as was demonstrated in 2001 with isolation of a genetically related H7N3 LPAIV strain in a cinnamon teal in Bolivia. However, the definitive source of the H7N3 Chilean LPAIV still remains unknown.     

Serum biochemical analytes in captive Amazonian manatees (Trichechus inunguis)

Background: Establishment of reference values for serum biochemical analytes is important for monitoring health and physiological status of captive animals. Objective: The purpose of this study was to measure and report ranges for serum biochemical analytes in Amazonian manatees (Trichechus inunguis). Methods: Blood samples were collected from 24 healthy captive Amazonian manatees that comprised a mixture of adults, subadults, and calves and males and females; serum analytes were measured and analyzed using a dry reagent bench‐top chemical analyzer. Comparisons were made between sexes and with previously published values of closely related species. Results: Medians and ranges (minimum–maximum) of values for the analytes were: lactate dehydrogenase (LDH), 151 (111–278) U/L (n=20); creatine kinase, 144 (76–315) U/L (n=11); alanine aminotransferase, 10 (2–28) U/L (n=18); aspartate aminotransferase, 14 (5–28) U/L (n=21); γ‐glutamyltransferase, 47 (36–73) U/L (n=21); amylase, 1428 (1010–1874) U/L (n=21); alkaline phosphatase, 73 (36–141) U/L (n=19); total protein, 6.8 (6.2–8.0) g/dL (n=24); albumin, 3.3 (2.6–4.1) g/dL (n=21); cholesterol, 188 (101–399) mg/dL (n=21); triglycerides, 126 (60–236) mg/dL (n=21); glucose, 47 (22–69) mg/dL (n=21); urea, 43 (21–69) mg/dL (n=21); uric acid, 1.1 (0.5–1.8) mg/dL (n=22); creatinine, 2.2 (1.5–3.3) mg/dL (n=22); total bilirubin, 0.2 (0.2–2.0) mg/dL (n=21); calcium, 12.7 (10.2–18.6) mg/dL (n=24); iron, 282 (207–457) μg/dL (n=13); and magnesium, 6.9 (4.3–8.9) mg/dL (n=20). With the exception of LDH, no differences were observed between sexes. Conclusions: The ranges obtained in this study provide important preliminary estimates for concentrations and activities of serum analytes in Amazonian manatees until a larger reference interval study can be conducted.     

Ovicidal activity of seven <Emphasis Type="Italic">Pochonia chlamydosporia</Emphasis> fungal isolates on <Emphasis Type="Italic">Ascaris suum</Emphasis> eggs

The ovicidal effect of the nematophagous fungus Pochonia chlamydosporia on eggs of Ascaris suum was tested under laboratory conditions. A. suum eggs were plated on 2% water–agar with seven fungal isolates (Isol. 5, Isol. 31, Isol. 1, VC1, Isol. 12, Isol. 22 and VC4) and control without fungus. After 5, 7, 10, 14, 15 and 21 days of incubation, approximately 100 eggs were removed from the plates and classified according to the following parameters: type 1, biochemical and physiological effect without morphological damage to the eggshell, type 2, lytic effect with morphological alteration of the eggshell and embryo and type 3, lytic effect with morphological alteration of eggshell and embryo showing hyphal penetration and internal egg colonization. The isolates effectively destroyed A. suum eggs and all types of effects were observed during the experiment. There was no variation in ovicidal capacity (type 3 effect) among the isolates (p > 0.05) throughout the experiment. After 21 days, isolate 5 showed the highest percentages of type 3 effect (58.33%). The results indicated that P. chlamydosporia (Isol. 5, Isol. 31, Isol. 1, VC1, Isol. 12, Isol. 22 and VC4) can destroy A. suum eggs and is, therefore, a potential biological control agent of nematodes.     

Seroprevalence of Toxoplasma gondii antibodies in captive elephants (Elephaus maximus maximus) in Sri Lanka

Serum samples collected during August 2003-June 2004 from 45 privately owned captive and 8 elephants from the Pinnawala Elephant Orphanage were tested for the presence of antibodies against Toxoplasma gondii using the direct modified agglutination test (MAT). Antibodies were found in sera of 14 of 45 (32%) privately owned elephants with titers of 1:25 in three, 1:50 in three, 1:100 in three, 1:200 in three, and 1:400 in three elephants. The elephants from Pinnawala Elephant Orphanage were seronegative. This is the first report of T. gondii seroprevalence in elephants in Sri Lanka.     

Microvascular development and growth of uterine tissue during the estrous cycle in mares

OBJECTIVE: To document uterine growth and microvascular development in the endometrium of uteri with differing degrees of fibrosis as well as uterine growth throughout the estrous cycle of mares. ANIMALS: 30 mares. PROCEDURE: Uterine tissue was obtained during the breeding season from a slaughter facility. Stage of estrous cycle of the mares was assessed on the basis of ovarian structures and plasma progesterone concentrations. Endometrium was characterized by use of light microscopy, and blood vessel walls were marked by histochemical techniques. Microvascular development was evaluated by a computerized image analysis system. Growth of uterine tissue was based on cellular content of DNA and RNA, RNA:DNA, and protein:DNA. RESULTS: Significant differences in vascular density were not observed in the endometrium of uteri obtained from mares euthanatized during the follicular or luteal phase of the estrous cycle, regardless of whether endometrial classification of degree of fibrosis was considered. There was a 3-fold increase in amount of DNA and RNA of endometrial cells in the follicular phase when compared to myometrium. Hypertrophy of endometrial tissue during the luteal phase was reflected by a significant increase in cell protein content and protein:DNA. CONCLUSIONS AND CLINICAL RELEVANCE: Endometrial growth of vascular tissues during the estrous cycle may be coordinated with development of nonvascular tissue. Estrogen and progesterone may play a role in regulation of uterine growth and angiogenesis.     

The Influence of Oxygen Tension on Cumulus Cell Viability of Canine COCs Matured in High-Glucose Medium

High in vitro oxygen (O₂) tensions are associated with enhanced levels of reactive oxygen species and cumulus oocyte complex (COC) apoptosis. The objective of this study was to determine the influence of O₂ tension on cumulus cell (CC) viability from canine oocytes. Cumulus oocyte complexes were distributed into three groups (CG, T20 and T5) and two O₂ tension levels (20% and 5%). The control group (CG) was matured in vitro in a humidified atmosphere with 5% CO₂ in air in TCM199 with 26.19 m m sodium bicarbonate, 10% (v/v) foetal calf serum (FCS), 0.10 m m gentamicin, 0.20 m m pyruvic acid, 20 μg/ml oestradiol, 0.5 μg/ml follicle-stimulating hormone, 0.03 IU/ml human chorionic gonadotropin, and 1.0 μg/ml human somatotropin. Groups T20 and T5 were matured under 20% or 5% O₂ tensions respectively in a high-glucose medium, without FCS. T20 and T5 were as CG, and supplemented with 0.1% Polyvinyl Alcohol, and 5.5 m m glucose. After 48 h of IVM, CCs from COCs were stained with propidium iodide (1.50 m m ). The results showed that viability of CCs (cytoplasmic features and nuclear morphological integrity) was different for the three groups. Rates of apoptosis were at 57.9% (521/900) for CG, 54.4% (490/900) for T20 and 38.9% (350/900) for T5 (p < 0.001). Predominant features in apoptotic cells (n = 1361) were DNA nuclear fragments (94.0%). It was concluded that CCs of canine COCs cultured in high-glucose medium showed significantly less apoptosis than those cultured in medium with FCS. Low O₂ tension was efficient in reducing apoptosis in canine CCs.     

Assessment of dexmedetomidine/ketamine anesthesia in golden-headed lion tamarins (Leontopithecus chrysomelas)

OBJECTIVE: To compare the anesthetic and cardiovascular effects of dexmedetomidine/ketamine combinations in golden-headed lion tamarins. STUDY DESIGN: Prospective blinded randomized study. ANIMALS: Six healthy golden-headed lion tamarins, three males and three females were studied (mean body weight +/- SD = 0.498 +/- 0.054 kg). METHODS: The animals were given one of the dexmedetomidine/ketamine combinations (0.005/10, 0.01/10, and 0.01/5 mg kg(-1) IM; treatments 5D/10K, 10D/10K, and 10D/5K, respectively) on three successive occasions. Time to sedation and recumbency, as well as anesthesia, standing, and walking times were recorded. Heart and respiratory rate, arterial blood pressure, hemoglobin saturation, and rectal temperature were recorded during anesthesia. Sedation, muscle relaxation, and auditory response were evaluated after drug administration. RESULTS: Heart rate decreased after two combinations (10D/5K and 10D/10K). Respiratory rate and rectal temperature progressively decreased in all the treatments. Arterial blood pressures were maintained in 5D/10K and 10D/5K treatments and decreased after 10D/10K administration. There were no differences in sedation and recumbency time between the treatments, but anesthesia time was significantly longer in treatment 10D/10K (67.80 +/- 13.30 minutes) compared to treatments 10D/5K (44.54 +/- 8.25 minutes) and 5D/10K (30.60 +/- 6.80 minutes). Standing time was shorter in treatment 10D/10K (11.13 +/- 10.01 minutes) than in treatments 5D/10K (18.20 +/- 10.03 minutes) and 10D/5K (19.12 +/- 14.55 minutes), and walking time was longer in treatment 5D/10K (20.00 +/- 12.46 minutes) compared with treatments 10D/10K (16.00 +/- 3.43 minutes) and 10D/5K (12.40 +/- 5.02 minutes). Anesthetic quality was significantly better 5 and 15 minutes after treatments 10D/10K and 10D/5K than after treatment 5D/10K. Analgesia scores were higher after administration of 10D/10K than after the 5D/10K treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Dexmedetomidine/ketamine is useful for chemical immobilization of golden-headed lion tamarins, but bradycardia and hypotension warrant close monitoring.     

Identification and quantification of benzimidazole resistance polymorphisms in Haemonchus contortus isolated in Northeastern Brazil

Haemonchus contortus is the most prevalent nematode in Brazil. The objective of this study was to select 6 populations of H. contortus of known or suspected benzimidazole resistance status and characterize these using quantitative real-time polymerase chain reaction (qPCR) for single nucleotide polymorphisms (SNPs) F200Y, F167Y and E198A in the β-tubulin isotype 1 gene. qPCR was performed using DNA from a pool of 10 adult male H. contortus from a single animal per farm. Faecal egg count reduction test (FECRT) and egg hatch test (EHT) were used to determine the resistance status. Samples were obtained from 6 farms located in 5 counties in the Ceará State: Tauá, Boa Viagem, Quixadá, Santa Quitéria and Solonópole. The inbred-susceptible-Edinburgh (ISE) isolate was used as reference for comparative purposes in the qPCR. Benzimidazole resistance was detected by FECRT on all farms with efficacy values ranging from 0 to 51%. EC50 values as determined by EHT were all above 1.49 μg/ml. High frequencies of the resistant SNPs F200Y and F167Y alleles were detected but no resistance was detected at SNP E198A. Our results suggest that the SNPs F167Y and F200Y are both important for benzimidazole resistance in the studied populations.