A survey of mutations in the Cochliomyia hominivorax (Diptera: Calliphoridae) esterase E3 gene associated with organophosphate resistance and the molecular identification of mutant alleles

Cochliomyia hominivorax (Calliphoridae) is one of the most important myiasis-causing flies and is responsible for severe economic losses to the livestock industry throughout the Neotropical region. In Brazil, C. hominivorax has been controlled mainly with organophosphate (OP) insecticides, although the inappropriate use of these chemicals can result in the selection of resistant flies. Changes in carboxylesterase activity have been associated with OP insecticides in some arthopodan species. In this work, we isolated and characterized part of the E3 gene in C. hominivorax (ChE7), which contained the same substitutions responsible for the acquisition of OP hydrolase activity in Lucilia cuprina (Calliphoridae). Digestion of the polymerase chain reaction products with a restriction enzyme that specifically recognized the mutation site unambiguously differentiated wild and mutated esterase alleles. The PCR-RFLP assay therefore provided a fast, reliable DNA-based method for identifying C. hominivorax individuals with a mutation in the esterase gene. Further bioassays to determine the association of this mutation with OP resistance in C. hominivorax should allow the development of more effective strategies for managing this species.     

Bluetongue in The Netherlands; description of the first clinical cases and differential diagnosis. Common symptoms just a little different and in too many herds

For the first time Bluetongue (BT) has been diagnosed in the Netherlands. The clinical symptoms of BT on five farms during the first outbreak ever in the Netherlands are described. Fever and swollen sensitive coronets leading to reluctance to stand and walk were sometimes the first symptoms. Later lesions in the mouth occurred with foamy salivation and respiratory problems. In other cases a swollen head with swollen lips and foamy salivation were the first clinical signs. Also sudden death occurred. In the first sixteen confirmed cases morbidity and mortality were lower than described in outbreaks in other countries. Good collaboration between practitioners, specialists of the Animal Health Service (GD-Deventer), and specialists of the Food and Consumer Product Safety Authority (VWA) and CIDC-Lelystad (Wageningen UR) led to a rapid notification and ultimately confirmation of the suspected diagnosis BT.     

The biochemical, morphological and virulence profiles of Bacillus anthracis isolated in the Kruger National Park

The biochemical, morphological and virulence profiles of 44 Bacillus anthracis isolates, obtained from various localities and carcass remains of wild animals in the Kruger National Park, South Africa, were examined. The morphological characteristics tested for included: the formation of capsules on bicarbonate agar, the motility of the vegetative organism, the presence of haemolysis on blood tryptose agar, the sensitivity of the vegetative organism to bacteriophage, the production of lecithinase on egg yolk agar, the liquefaction (hydrolysis) of gelatine and the capability of each isolate to produce mucoid colonies when grown on bicarbonate agar with horse serum in an atmosphere containing CO2. The API 50CHB and 20E systems were used to evaluate the biochemical activity of each isolate. The virulence of each isolate was determined by its LD50, using an inbred line of Balb/C mice. A clear pattern in the biochemical reactions emerged that appeared to be specific for each isolate. On the API 50CHB test strip, only 2 of the 44 isolates gave a 100% positive reaction to all 10 of the biochemical substances to which it was supposed to react, 9 gave positive results to 90%, 19 were positive to 80%, and 14 were positive to 70%. The reactions on the API 20E were completely different from what was expected, with only 1 of the biochemical activities (gelatinase production) showing a positive reaction to all the isolates. The virulence test indicated that 27/44 isolates could be regarded as highly virulent with a LD50 of less than 1,000 organisms, and the rest of the isolates as virulent with a LD 50 of 1,001-10,000 organisms.(ABSTRACT TRUNCATED AT 250 WORDS)     

Presence of fluoroquinolone-resistant coliforms in poultry litter

Litter was collected from four turkey farms (eight houses) with a history of fluoroquinolone (FQ) treatment failure, 10 adult broiler breeder chicken farms (43 houses) with one having a history of FQ treatment, and 30 broiler chicken farms (110 houses) with 24 having a history of FQ treatment. In the turkey litter, the percentage of nalidixic acid-resistant (at 100 microg/ml) coliforms/total number of coliforms ranged from 0.6% to 61.9%. Two of the four farms had houses containing coliforms resistant to the two FQs, enrofloxacin (1 microg/ml) and sarafloxacin (1 microg/ml). There was also multiple resistance to other antimicrobials on all four turkey farms (ampicillin, tetracycline, chloramphenicol, kanamycin). The level of total coliforms from the adult broiler breeder litter was low, and there were no nalidixic acid-resistant isolates from any of the 10 farms. In the broiler chickens, 7 of 91 houses with a history of FQ usage contained coliforms resistant to nalidixic acid; however, 2 of the 19 houses on farms with no history of FQ usage had nalidixic acid-resistant coliforms. All of the broiler farms with nalidixic acid-resistant isolates were also resistant to the FQ sarafloxacin, whereas only 3 of the 24 treatment history farms and 1 of the no-treatment history farms exhibited enrofloxacin-resistant coliforms in the litter.     

Classical swine fever in The Netherlands 1997-1998: a description of organisation and measures to eradicate the disease

The central and regional organisation of the campaign to eradicate the CSF epidemic in the Netherlands in 1997/1998 is described. The main instruments used in the campaign were based on stamping-out and movement restrictions specified by the European Union. Additional instruments were used for the first time, namely, pre-emptive culling of contact and neighbouring farms, compartmentalisation of transport, monthly serological screening in established surveillance areas and supervised repopulation of all farms in the former surveillance zone. Two other measures, the killing of very young piglets and a breeding ban were introduced to reduce production in established surveillance zones. Several factors complicated the eradication campaign, for instance, the late detection of the first infection; artificial insemination as a source of infection; the organisation of pig farming in the Netherlands, with its highly concentrated production and dependence on the transport of stock from one unit to another; insufficient rendering capacity; decreasing sensitivity of clinical inspection; and extremely high costs.     

Detection of anti-Leishmania infantum antibodies in sylvatic lagomorphs from an epidemic area of Madrid using the indirect immunofluorescence antibody test

An outbreak of human leishmaniosis was confirmed in the southwest of the province of Madrid, Spain, between July 2009 and December 2012. Incidence of Leishmania infection in dogs was unchanged in this period, prompting a search for alternative sylvatic infection reservoirs. We evaluated exposure to Leishmania in serum samples from animals in the area with an indirect immunofluorescence test (IFAT). Using promastigotes from six culture passages and a 1/25 threshold titer, we found anti-Leishmania infantum seroreactivity in 9.3% of cats (4 of 43), 45.7% of rabbits (16/35) and 74.1% of hares (63/85). Use of promastigotes from >10 in vitro passages resulted in a notably IFAT lower titer, suggesting antigenic changes during extended culture. Postmortem inspection of seropositive animals showed no clinical signs of infection. The results clearly suggest that asymptomatic hares were the main reservoir in the outbreak, and corroborate IFAT as a sensitive serological surveillance method to detect such cryptic Leishmania infections.     

Increased renal vascular resistance in dogs with hepatic disease

Doppler ultrasound is a non-invasive technique that can be used to estimate vascular resistance by calculation of resistive index (RI) and pulsatility index (PI). Liver disease may increase renal RI and PI, and in humans with liver disease the indices are monitored to attain prognostic information. Systemic hypertension has been found in dogs with hepatic disease and is also related to increased renal vascular resistance in humans. The aim of this study was to examine renal vascular resistance increases in dogs with hepatic disease and to ascertain whether these may be related to blood pressure increases and biochemical parameters. Twenty dogs with hepatic disease were evaluated. The mean renal RI, PI, and systolic blood pressure were significantly higher than in normal animals. A positive correlation was found between the indices and alkaline phosphatase but not with systolic blood pressure. It is concluded that renal vascular resistance may increase in dogs with hepatic disease and in this study was above the limit value in 50% of the animals.     

Effects of dobutamine on cardiac index and arterial blood pressure in isoflurane-anaesthetized horses under clinical conditions

Volatile agent-induced hypotension may contribute to anaesthetic-related morbidity and mortality in horses. Dobutamine is commonly used to support arterial blood pressure (ABP) but little is known about its cardiovascular effects under clinical conditions. The aim of this clinical study was to elucidate the relationship between cardiovascular function and dobutamine infusion in isoflurane-anaesthetized horses. Forty-four horses anaesthetized for a variety of surgical procedures were studied. Premedication with acepromazine, methadone and detomidine was followed by induction of anaesthesia with ketamine and midazolam. Anaesthesia was maintained with isoflurane vaporized in oxygen. Routine anaesthetic monitoring was applied and cardiac output was measured by lithium dilution. Dobutamine was infused to maintain mean ABP above 70 mmHg. The relationship between dobutamine infusion rate, heart rate (HR), ABP and cardiac index was investigated immediately prior to ( T ₀) and 15 min ( T ₁) after dobutamine infusion started, followed at 30 min intervals ( T ₂, etc.). Arterial blood pressure increased significantly after dobutamine infusion started, HR and cardiac index increased significantly only with dobutamine infusion in combination with surgical stimulus. Although isoflurane decreases blood pressure mainly by vasodilation, dobutamine is an effective treatment for hypotension under clinical conditions in isoflurane-anaesthetized horses. The effect of dobutamine is not directly proportional to dose and surgical stimulus probably contributes to the cardiovascular improvement.     

A retrospective analysis of radiation therapy for the treatment of feline vaccine‐associated sarcoma*

We retrospectively evaluated predictive prognostic factors in 73 cats with vaccine‐associated sarcoma given postsurgical curative (n = 46, most with clean margins) or coarse fractionated radiotherapy (n = 27, most with either macroscopic disease or dirty margins). The former animals displayed a median survival of 43 months and a median progression free interval (PFI) of 37 months, the latter reached a median survival of 24 months and a median PFI of 10 months. In cats undergoing coarse fractionated therapy, factors predictive of a better outcome included lack of visible mass (n = 10) as opposed to macroscopic disease (n = 17, survival: 30 versus 7 months, P = 0.025; PFI: 20 versus 4 months, P = 0.01), adjuvant chemotherapy for gross disease (n = 5/17, survival: 29 versus 5 months, P = 0.04) and a smaller number of surgeries preceding radiation therapy (coeff = 0.41, P = 0.03). The Ki67 index was not predictive for survival. We concluded that postsurgical curative and coarse fractionated radiotherapy are effective legitimate options for managing vaccine‐associated sarcomas.