In vitro inactivation of two Egyptian A/H5N1 viruses by four commercial chemical disinfectants.

The highly pathogenic H5N1 avian influenza virus (A/H5N1) devastated the poultry industry and posed a serious health threat. Cleaning and disinfection are essential parts of preventative and postoutbreak management of A/H5N1 infections in poultry. In this preliminary study, we used suspension and carrier tests to evaluate the impact of concentration, time of exposure, surface porosity, and organic matter on the ability of four commercial chemical disinfectants to inactivate two A/H5N1 viruses of clade 2.2.1 isolated in 2006 and 2010 from broiler flocks in Egypt. Viruses were incubated with 0.5%, 1%, and 2% of formalin, glutaraldehyde, TH4, and Virkon S for 15, 30, 60, and 120 min at room temperature (22 +/- 2 C). In suspension tests, in the absence of organic matter, all disinfectants, at each concentration, except Virkon S 0.5%, effectively inactivated virus suspensions after a 15-min exposure time. In the presence of organic matter, the use of low concentrations of formalin (0.5%), glutaraldehyde (0.5%), or Virkon S (0.5%) was not sufficient to inactivate the viruses after 15 min. In gauze carrier tests, only formalin at any concentration for 15 min was sufficient to inactivate the viruses, whereas different concentrations or exposure times were required for glutaraldehyde (0.5% for 60 min), TH4 (0.5% for 30 min), and Virkon S (0.5% for 60 min or 1% for 30 min). In wood carrier tests, total inactivation of the virus was obtained at concentrations of 0.5% for 30 min (formalin and TH4) or 60 min (glutaraldehyde and Virkon S). This study emphasizes the need to use high concentrations of and/or extended time of exposure to disinfectants for efficient inactivation of A/H5N1, particularly in the presence of organic matter or different surfaces, which are common in poultry operations. In addition, it seemed that the virus isolated in 2010 was more resistant to disinfectants than the isolate from 2006 when wood was used as a carrier.

Campylobacter in chicken carcasses and slaughterhouses in Malaysia.

This study was conducted to determine the Campylobacter contamination rate of chicken carcasses and the processing lines of modern processing plants in Malaysia. Three hundred sixty samples were collected from 24 flocks of broiler chickens at 12 modern poultry processing plants in 6 states of Malaysia. Fresh fecal droppings were collected from crates in the arrival area. Neck skin samples were taken from processed chicken carcasses at 3 different processing stages: before inside-outside washing, after inside-outside washing and post chilling. Swab samples from the scalding tank, chilling tank and conveyer belt before chilling were also collected to determine contamination with Campylobacter in the slaughter house environment prior to slaughter. Isolation for Campylobacter was performed following ISO 10272-1:2006(E). The overall of contamination rate with Campylobacter at the 12 plants was 61.0% (220/360). Eighty point six percent of the samples from before the inside-outside wishing step were contaminated with Campylobacter, as were 62.5% of the samples after the inside washing and 38.9% after the post-chilling step. This study shows extensive contamination of chicken carcasses and slaughtering houses in Malaysia with Campylobacter.

A questionnaire survey on helminth control on horse farms in Brandenburg, Germany and the assessment of risks caused by different kinds of management.

Control strategies for horse helminths are controversially discussed, and no ideal strategy exists. Presently, the spreading of anthelmintic resistance against all three classes of anthelmintics is of special concern. Advice on how to slow the development of anthelmintic resistance does not seem to have reached the majority of horse owners yet. In our study, we wanted to capture the current standard of helminth control and to analyse the effectiveness of these control strategies. Seven hundred horse farms in Brandenburg, Germany were sent a questionnaire in June/July 2006 asking various questions on farm structure and practices of helminth control. Two hundred thirty-five farms (33.6%) with 6,007 horses in total returned the questionnaire. The number of horses held on each farm varied from 1 to 320. From those returned questionnaires, a random sample of 126 horse farms and 1,407 horses was selected for faecal examination from August to December 2006. The questionnaire results from these farms were correlated with results from the faecal examination to perform risk analysis via multivariate regression. For each farm, results from faecal examination were summarized in index values. Risk was defined as an over-average level of strongyle egg shedding. Risk factors as established by multivariate regression analyses were integrated into three models. Rare deworming and infrequent cleaning of stables were identified as significant risk factors throughout all three models. Additional risk factors could be identified with respect to individual models: elevated percentage of young horses on a farm, not using macrocyclic lactones in the deworming process and dosing anthelmintics on the basis of weight and height. Pasture hygiene did not lead to significant risk reduction, indicating that such measures are inadequately performed on many farms. The questionnaire on its own gave us an insight into control strategies throughout a representative scope of farms in Brandenburg, Germany: faeces were collected from pasture on 22% of horse farms, 79% cleaned out the stables daily. A number of other measures were employed. With respect to anthelmintic use, 21% of the farms stated to use results from faecal examination as a criterion for deworming. We could also identify the most common deworming frequencies, with adult horses being dewormed two times a year (46% of farms) and foals being dewormed four times a year (35% of farms). The frequency ranged from 1 to 7 and 2 to 12 times, respectively.

Prevalence of helminths in horses in the state of Brandenburg, Germany.

The objective of the study was to estimate the prevalence of helminths in the horse population of the state of Brandenburg, Germany. One hundred and twenty-six horse farms in the state were selected by randomised stratified sampling. In total, 1,407 horses across all farms were examined coproscopically. The experimental unit was the horse farm: a farm was considered infected when at least one horse on the farm investigated was positive for helminth eggs. Animal details such as age, breed and sex were collected for all study horses and analysed for risk of infection. Risk was defined as horses having an above-average shedding of strongyle eggs. The following prevalence on horse farm level were established: Cyathostominae (98.4%), ascarids (16.7%), tapeworms (14.3%), pinworms (8.7%) and strongyloides (4,0%). The large strongyle Strongylus vulgaris was identified on only one farm. Liver flukes and lungworms were not found. Age, breed and sex were identified as risk factors for high shedding of strongyle eggs of individual animals: odds ratios for higher shedding intensities were 4.18 for yearlings and 2.42 for fillies compared to adult animals, and 3.69 for heavy breeds and 4.94 for wild horses compared to thoroughbreds. Mares and stallions did shed more strongyle eggs than geldings. Knowledge about the helminth prevalence will allow the issuance of specific treatment recommendations. Furthermore, the information on risk factors of individual horses will facilitate targeting single animals for selective treatments.

A cross-sectional study of Salmonella in pork products in Chiang Mai, Thailand.

The occurrence of Salmonella in food of animal origin in Chiang Mai province was investigated by using a cross-sectional study during several phases of the pork production chain (cutting, transport, and retail) and of the environment in the cutting unit of a slaughterhouse. In total, 173 pork samples were obtained during the cutting phase, 173 samples from transported pork, 200 samples from retail products, and 300 samples from the slaughterhouse environment. Salmonella was detected in 55.5% of freshly cut pork, 70.5% of transported pork, and 34.5% of retail products. The five most prevalent Salmonella serotypes identified were Rissen (45.3%), Typhimurium (16.3%), Krefeld (10.6%), Stanley (6.3%), and Lagos (6.0%). Carcass contamination prior to cutting and in the slaughterhouse environment appeared to be important sources of Salmonella in transported pork and retail products. As Salmonella was also found during early stages of the slaughter process, attention should focus on all stages of the pork production chain to reduce contamination level and consumer risk of infection.

Evaluation of medicinal plants from Mali for their in vitro and in vivo trypanocidal activity.

Water, methanol and dichloromethane extracts prepared from various parts of 40 medicinal plant species from Mali were investigated for their trypanocidal activity against Trypanosoma brucei brucei. Of a total of 165 extracts tested in vitro in the Low Inoculation Long Incubation Test (LILIT), 24 extracts showed a high trypanocidal activity. Using the Long-Term Viability Assay (LtVA) for corroboration of the results of the 24 extracts, it was found that 15 samples had minimum inhibitory concentration (MIC) values > 10 microg/ml, eight MIC values of 100 microg/ml and one MIC values of 50-100 microg/ml. So far, four extracts with MIC values < or = 100 microg/ml were tested for antitrypanosomal activity in mice, experimentally infected with Trypanosoma brucei brucei. Only, the aqueous extracts of the leaves of Terminalia avicennioides Guill. and Perr. (Combretaceae) and the stem bark of Ceiba pentandra (L.) Gaertn. (Bombacaceae) were able to reduce the parasitaemia in animals treated at the dose of 100 mg/kg b.w. (intraperitoneally, two times daily for 3 days) and of 150 mg/kg b.w. (per os, two times daily for 3 days), respectively. The reduction of parasitaemia was, however, statistically significant (p=0.002) only in case of treatment with Terminalia avicennioides.

Microbial contamination of pig carcasses at a slaughterhouse in Vientiane capital, Lao PDR.

A cross-sectional study was conducted to determine microbial contamination of pig carcasses at a slaughterhouse in Vientiane, capital of Lao People Democratic Republic (Lao PDR). Between November 2004 and April 2005, 62 pig carcasses were randomly selected. From each carcass, pooled swabs (from "1" prior to and "2" after evisceration) and 25 g of tissue of mesenteric lymph nodes (MLN) were collected. The swab samples were examined for Aerobic Plate Count (APC) and Enterobacteriaceae Counts (EBC) and cultured for Salmonella. The lymph nodes were cultured for Salmonella only. Swabs1 and 2 had mean APC of 4.70 and 4.85 log10CFU/cm2, respectively. These two means were significantly (p = 0.0001) different. The means of EBC were 2.81 log10CFU/cm2 for Swab 1, and 2.98 log10CFU/cm2 for Swab 2. The difference were also statistical significant (p = 0.0001). The frequency of Salmonella isolation from Swab 1 was 46.8%, for Swab 2 was 66.1%, and from mesenteric lymphnodes was 53.2%. Eight different Salmonella serotypes were identified. The most frequent (29.1%) serotype was S. Rissen, followed by S. Anatum (26.2%), S. Derby (18.4%), and S. Elisabethville (8.7%). The other serotypes identified were S. Amsterdam (7.8%), S. Typhimurium (4.9%), S. Agona (2.9%), and S. Enteritidis (1.9%). Results of this study showed the levels of contamination with aerobic bacteria and Enterobacteriaceae were higher than recommended standards, and the carcasses were contaminated with Salmonella.

Salmonella serovars along two beef chains in Ethiopia.

INTRODUCTION: Salmonella has been reported from foods and the food production environment, with outbreaks occurring in the human population worldwide. METHODOLOGY: A survey on Salmonella in two beef production lines (a beef abattoir line and a processing line) in Addis Ababa, Ethiopia was conducted, with a total of 668 various samples randomly collected from animal-related materials, the environment, and a beef product (mortadella). RESULTS: Overall, a 12.9% prevalence (26.3% from the abattoir line, 5.3% from the processing plant line) was observed. The prevalence in the abattoir line environment (36.6%) was higher than that in animal-related samples (14.7%); the reverse was true for the processing plant line. Out of 86 isolates, 10 serovars were identified, and 8 remained unidentified. The predominant serotypes were S. Saintpaul (32.5%), S. Muenchen (19.8%), and S. Larochelle (12.8%). S. Kastrup and S. London were isolated for the first time in Ethiopia. CONCLUSIONS: Data indicate open ports of entry for Salmonella, with possible transfer along the line. Further investigations from farm to fork are recommended in order to identify these positions of entry.

Application of field methods to assess isometamidium resistance of trypanosomes in cattle in western Ethiopia.

This study assessed the degree of isometamidium resistance of trypanosomes infecting cattle in the upper Didessa valley of western Ethiopia. An initial prevalence study was conducted to identify sites with a high risk of trypanosmosis in cattle. The trypanosome prevalence varied widely, with two sites, Kone (21.3%) and Village 1 settlement (15%) having a relatively high prevalence based on the phase-contrast buffy-coat technique (BCT). In the highest risk area, the Kone settlement, an isometamidium block treatment study was conducted from April to June 2001. A total of 300 cattle were included in this study, 100 from each of three villages (Cheleleki, Kolu and Burka). At day minus 14 of the study, all 300 cattle were treated with diminazene aceturate at 7 mg/kg body weight. Subsequently, these cattle were ear-tagged and randomly assigned into two groups, 50 controls and 50 for isometamidium treatment in each village. Fourteen days later (day 0), the 50 treatment cattle were given isometamidium chloride at 1 mg/kg body weight. Both groups of cattle were then examined for trypanosome parasites using BCT every 14 days until 84 days. The two indices used in assessing isometamidium resistance, namely the proportion of infections during an 8-week follow-up period and the ratio of mean hazards in an isometamidium treated versus untreated group, provided consistent results across the three villages. In Burka village, both indices demonstrated the presence of isometamidium resistance trypanosome infections while in Cheleleki and Kolu villages, both indices did not indicate significant levels of resistance. There were significant differences between the Kaplan-Meier survival estimates of the control and treatment groups in Cheleleki (P < 0.01) and Kolu (P < 0.05) but not in Burka (P > 0.05).

A field study to estimate the prevalence of Trypanosoma equiperdum in Mongolian horses.

From May to July 2000, a cross-sectional study was conducted to estimate the prevalence of Trypanosoma equiperdum in the horse population of the central province (Tuv aimag) of Mongolia. On average, four herds were selected from each of the 29 aimag subdivisions (119 herds). From each herd, 10 horses were sampled in proportion to sex and age categories in the respective herds (1190 horses). Sera from 1122 horses were analysed for T. equiperdum antibodies using two serological assays, the complement fixation test (CFT) and the enzyme-linked immunosorbent assay (ELISA). The crude estimate of the CFT and the ELISA seroprevalence was 7.6 and 6.7%, respectively. Concordance between the CFT and ELISA results was high (96%). The highest number of CFT positive animals was detected in one herd in Möngönmorit (6/10), followed by herds in Bayandelger (5/10) and in Bayantsagaan (5/10). Poor body condition was significantly correlated with positive serological status in both CFT and ELISA. A history of abortion appeared to be a risk factor for both CFT and ELISA seropositivity. Blood samples of all horses belonging to herds with at least three (3/10) seropositive animals (CFT and/or ELISA) were analysed by light microscopy and by PCR using a Trypanosoma (Trypanozoon) brucei specific primer pair. No trypanosomes or any other haemoparasites could be detected in Giemsa stained thin blood smears. Eight out of the 130 samples (6.2%) analysed by PCR gave positive signals. Seven out of the eight PCR positive horses were also serologically positive. One PCR (and ELISA) positive stallion from Möngönmorit showed emaciation, scrotal and preputial oedema and an oedematous skin plaque. From the serological and DNA-based results it is concluded, that trypanosome infections occur in horses in the Tuv aimag of Mongolia. Since at present neither serological nor DNA-based tests allow a subspecies specific identification within the subgenus Trypanozoon, no definitive diagnosis can be given for T. equiperdum. Whether the examined herds are infected with T. equiperdum or with T. evansi, the causative agent of surra, remains an open question. However, based on the clinical findings, the negative parasitological results and the concentration of conspicuous seroprevalences in single herds, circumstantial evidence supports the existence of infections with the causative agent of dourine.

Comparison of multilocus sequence typing (MLST) and repetitive sequence-based PCR (rep-PCR) fingerprinting for differentiation of Campylobacter jejuni isolated from broiler in Chiang Mai, Thailand.

We compared rapid fingerprinting using repetitive sequence-based PCR (rep-PCR) for subtyping Campylobacter jejuni isolates to the widely used multilocus sequence typing (MLST). Representative C. jejuni isolates (n = 16) from broilers were analyzed using MLST and rep-PCR. Both techniques demonstrated an equal discriminatory power of 0.8917, and 9 subgroups were identified. Clonal identification of all 16 isolates was identical for both techniques. The rep-PCR as described in this study may be used as a rapid and cost-effective alternative for subtyping of C. jejuni isolates, or as an effective screening tool in large epidemiological studies.

Comparison of Campylobacter contamination levels on chicken carcasses between modern and traditional types of slaughtering facilities in Malaysia.

A total of 360 samples including fresh fecal droppings, neck skins, and swab samples was collected from 24 broiler flocks and processed by 12 modern processing plants in 6 states in Malaysia. Ninety samples from 10 traditional wet markets located in the same states as modern processing plants were also collected. Microbiological isolation for Campylobacter was performed following ISO 10272-1:2006 (E). The overall rate of contamination for Campylobacter in modern processing plants and in traditional wet markets was 61.1% (220/360) and 85.6% (77/90), respectively. Campylobacter jejuni was detected as the majority with approximately 70% for both facilities. In the modern processing plants, the contamination rate for Campylobacter gradually declined from 80.6% before the inside-outside washing to 62.5% after inside-outside washing and to 38.9% after the post chilling step. The contamination rate for Campylobacter from processed chicken neck skin in traditional wet markets (93.3%) was significantly (P<0.01) higher than in modern processing plants (38.9%).

Prevalence and molecular characteristics of Vibrio spp. isolated from preharvest shrimp of the North Western Province of Sri Lanka.

This study investigated the prevalence and molecular characteristics of Vibrio spp. in farmed shrimp (Penaeus monodon) in Sri Lanka. A total of 170 shrimp samples (100 g of whole shrimp each) taken from individual ponds from 54 farms were collected 1 week prior to harvest from the North Western Province of Sri Lanka. Overall, 98.1% of the farms and 95.1% of the ponds were positive for Vibrio spp. in shrimp; at the pond level, V. parahaemolyticus (91.2%) was most common, followed by V. alginolyticus (18.8%), V. cholerae non-O1/non-O139 (4.1%), and V. vulnificus (2.4%). Multiple Vibrio spp. were detected in 20.6% of the ponds. None of the V. parahaemolyticus isolates (n = 419) were positive for the virulence-associated tdh (thermostable direct hemolysin) and trh (TDH-related hemolysin) genes. V. cholerae was confirmed by the presence of ompW, and all isolates (n = 8) were negative for the cholera toxin (ctxA) gene. V. cholerae isolates were serogrouped by PCR and identified as V. cholerae non-O1/non-O139. All four V. vulnificus strains, isolated from different ponds of two geographical regions, showed pathogenic potential; they belonged to vcgC sequence type, type B 16S rRNA genotype and contained a pilF polymorphism associated with human pathogenicity. The results of this study revealed the ubiquitous nature of vibrios in farmed shrimp. To minimize the potential risk of Vibrio infections due to handling or consumption of raw or undercooked seafood products, good manufacturing practices as well as proper handling and processing should be addressed.

Detection of multiple drug-resistant Trypanosoma congolense populations in village cattle of south-east Mali.

BACKGROUND: Tsetse fly-transmitted African animal trypanosomosis causes annual losses that run into billions of dollars. The disease is assumed to cause hunger and poverty in most sub-Saharan countries since it represents a serious impediment to sustainable livestock production. Both a cross-sectional and a longitudinal study were carried out from November to December 2007 to evaluate trypanosomosis risk and susceptibility of trypanosomes to trypanocidal drug treatment in village cattle populations in south-east Mali. METHODS: Eight purposively selected villages participated in the study. In each village, eight traps deployed along drainage lines over 24 hour duration were used to catch tsetse. One hundred systematically selected cattle in the study villages were examined for trypanosomes. All trypanosome-positive cattle were randomly allocated into two treatment groups: a group treated with 0.5 mg/kg bw. isometamidium chloride (ISMM) and a group treated with 3.5 mg/kg bw. diminazene aceturate (DIM). The cattle were monitored for trypanosomes at day 14 and 28 post-treatment. RESULTS: Of the 796 cattle examined, 125 (15.7%) were trypanosome-positive. Village trypanosome prevalences ranged between 11% and 19%. There were no significant (p > 0.05) differences in the village trypanosome prevalences. Trypanosoma congolense was the dominant trypanosome species accounting for 73% (91/125) of the infections and T. vivax the remainder. Twenty (31.7%) of the 63 cattle on 0.5 mg/kg bw. ISMM treatment were still positive14 days post-treatment. Of the 43 aparasitaemic cattle monitored to day 28, 25.6% (11) became parasitaemic, resulting in a cumulative failure rate of 49.2% (31/63). Trypanosoma congolense accounted for 77.4% (24/31) of failed ISMM treatments. The 62 cattle treated with 3.5 mg/kg bw. DIM resulted in 30.6% (19/62) failed treatments. Although 42.2% (19/45) of T. congolense positive cattle did not respond to DIM treatment, all T. vivax positive cattle responded positively to DIM treatment. CONCLUSIONS: The overreliance on trypanocides in the control of trypanosomosis will ultimately lead to multiple drug-resistant trypanosome populations as detected in villages in south-east Mali rendering the use of drugs doubtful. Effective alternative methods for trypanosomosis control ought to substitute chemotherapy to ensure sustainable cattle production in these villages. Since there is no single strategy for containing trypanocidal drug resistance, promotion of an integrated approach combining proven trypanosomosis control approaches in high trypanosomosis risk areas is most desirous. The best-bet strategy this study recommended for areas with multiple drug resistance included area-wide community tsetse control, control of co-infections to exploit self-cure against resistant trypanosome populations and the rational use of trypanocidal drugs which should be urgently promoted at all levels as a way of containing or reversing resistance.

Best-bet integrated strategies for containing drug-resistant trypanosomes in cattle.

BACKGROUND: African animal trypanosomosis is a major constraint to the rearing of productive livestock in the sub-humid Sudan-Sahel zone of West Africa where cotton is grown. Trypanosomosis is mainly controlled using trypanocidal drugs, but the effective use of drugs is threatened by the development of widespread resistance. This study tested integrated best-bet strategies for containment and/ or reversal of trypanocide resistance in villages in south-east Mali where resistance has been reported. METHODS: Four sentinel villages each from an intervention area (along the road from Mali to Burkina Faso) and a control area (along the road from Mali to Côte d'Ivoire) were selected for the study. Tsetse control was based on deltamethrin-treated stationary attractive devices and targeted cattle spraying between March 2008 and November 2009. Trypanosome-positive cattle were selectively treated with 3.5 mg/kg diminazene aceturate. Strategic helminth control using 10 mg/kg albendazole was also undertaken. During the intervention, tsetse densities along drainage lines, trypanosome infections and faecal egg counts in risk cattle (3 to 12 months of age) were monitored. RESULTS: Catch reductions of 66.5 % in Glossina palpalis gambiensis and 90 % in G. tachinoides were observed in the intervention area. Trypanosome prevalence was significantly (p < 0.05) lower in the intervention area (2.3 %; 1.3-3.6 %) compared to the control area (17.3 %; 14.8-20.1 %). Albendazole treatment resulted in a faecal egg count reduction of 55.6 % and reduced trypanosome infection risk (2.9 times lower than in the placebo group) although not significantly (p > 0.05). Further studies are required before confirming the existence of albendazole resistant strongyles in the study area. CONCLUSION: Integration of best-bet strategies in areas of multiple drug-resistance is expected to reduce trypanosome infection risk thus contributing to containment of trypanocidal drug resistance. Integrated best-bet strategies could therefore be considered a viable trypanosomosis control option especially in areas where multiple drug-resistance has been reported.

Detection of patent infections of Echinococcus granulosus ("sheep-strain", G1) in naturally infected dogs in Kosovo.

A survey was carried out to assess the occurrence of canine echinococcosis in naturally infected dogs in Kosovo. Using the flotation-ovassay technique, taeniid eggs were found in 23 (7.5%) out of a total of 305 dogs. Eggs from other helminths were detected as well: hookworms 139 (45.5%), Trichuris sp. 87 (28.5%), Toxocara sp. 42 (13.7%), Toxascaris leonina 21 (6.8%) and Dipylidium caninum eight (2.6%). From 21 of the 305 samples (6.9%), taeniids eggs could be collected. Using PCR primers specific for Echinococcus granulosus ("sheep strain", G1), four of these samples (1.3%) resulted positive. The E. granulosus isolates originated from each one stray dog, hunting dog, sheepdog and pet dog. A semi-quantitative analysis showed low to moderate egg counts (2-10 per 1 g faeces) in dogs positive for E. granulosus ("sheep strain", G1) whereas specimens with high (11-20) or very high numbers (> 20) of taeniid eggs were negative in the E. granulosus PCR. Using specific primers for the detection of E. multilocularis, all samples containing taeniid eggs were negative. This is the first report on identification of E. granulosus in dogs from Kosovo where human cystic echinococcosis is a significant medical problem.

Host preference of tsetse: an important tool to appraise the Nagana risk of cattle in the cotton zone of Mali.

Nagana, a vector-borne epizootic caused by trypanosomes, severely constrains the use of draught animals in the cotton zone of south-eastern Mali. The disease causes considerable economic losses for the local farmers due to high mortality and morbidity ensuing productivity losses. Nagana is routinely controlled by the use of trypanocides and an overreliance on their use throughout past decades resulted in multiple drug resistance of trypanosomes in most parts of West Africa's cotton belt. Designing alternative, effective vector control strategies requires an identification of the preferred hosts of tsetse flies through blood meal analysis as a prerequisite for estimating infection risk. A survey was, therefore, conducted between November 2008 and April 2009, catching 474 Glossina species which were dissected. Blood meals were smeared on filter paper (Whatman(®)-FTA-Cards) for laboratory analysis. DNA extractions and amplification using universal vertebrate cytochrome b primers of 120 assorted samples detected 74 DNA-containing specimens. The subsequent use of cattle-specific primers yielded 52 visible amplicons in the gel electrophoresis. Sequencing and BLASTN(®) analysis of the remaining samples revealed 19 blood meals matching with existing sequences of the human genome in Genbank(®). Two samples originated from crocodiles whereas one was unidentifiable.

Prevalence of Salmonella in marketed Penaeus monodon shrimps in North Western Province, Sri Lanka.

The examination of a total of 180 samples from both captured and cultured shrimps from Sri Lanka for the presence of Salmonella revealed an overall prevalence of 12.8%. The prevalence of Salmonella in captured shrimps and cultured shrimps was 14.4% and 11.1% respectively, but thedifference was not statistically significant (p = 0.66). The serovar most frequently isolated was S. Newport (47.8% of the isolates), followed by S. Weltevreden (8.7%).

Rapid identification of isometamidium-resistant stocks of Trypanosoma b. brucei by PCR-RFLP.

Analyses were made on the adenosine transporter-1 gene in Trypanosoma brucei (TbAT1), encoding a P2-like nucleoside transporter, from T. brucei brucei field stocks to investigate a possible link between the presence of mutations in this gene and isometamidium resistance. We have analysed the gene from 11 isometamidium-sensitive field stocks isolated from cattle in Uganda, two sensitive reference clones and two resistant reference clones. A sequence alignment showed that the isometamidium-sensitive T. b. brucei contained the wild-type sequence patterns. In contrast, the isometamidium-resistant T. b. brucei stocks showed the mutant-type sequence patterns with six point mutations that had previously been reported in a laboratory-derived arsenical-resistant T. brucei strain. To analyse the restriction fragment length polymorphism pattern of a fragment of TbAT1 (nucleotides 430-1108), the 677-bp polymerase chain reaction products from eight of the isometamidium-sensitive and two of the isometamidium-resistant T. b. brucei were subjected to digestion with Sfa NI. The results revealed two different banding patterns: the digest resulted in fragment sizes of 566 and 111 bp in the case of TbAT1 from isometamidium-sensitive stocks, whereas it produced fragment sizes of 435 and 242 bp in the case of TbAT1 from isometamidium-resistant stocks. Thus, the isometamidium-sensitive and isometamidium-resistant T. b. brucei could be successfully distinguished by digestion with the restriction endonuclease Sfa NI.