Cryptosporidium and Giardia infections in dairy calves in southern Ethiopia.

Giardia and Cryptosporidium are the most common enteric protozoan parasites causing diarrhea in humans and animals worldwide. This study was conducted with the objectives of estimating prevalence and identifying risk factors for Cryptosporidium and Giardia infections in dairy calves in selected districts of southern Ethiopia. Fecal samples (n = 330) were collected from calves in 92 farms. The monoclonal antibody-based commercial direct immunofluorescent kit was used to test the samples for Cryptosporidium oocysts and Giardia cysts. A questionnaire survey was also administered to collect data on potential risk factors of infections. The results showed a farm-level prevalence of 69.6% (95% confidence interval [CI]: 59.1-78.7%) for Cryptosporidium and 38.04% (95% CI: 28.1-48.8%) for Giardia. Likewise, an overall animal level prevalence of 13.0% (95% CI: 9.6-17.2%) for Cryptosporidium and 9.7% (95% CI: 6.7-13.4%) for Giardia was found. At the farm level, multivariate logistic regression model showed that calves in smallholder farms were 5.3 times more likely to shed Cryptosporidium oocysts than calves in commercial farms (p=0.019). However, in case of Giardia, calves in commercial farms were 5.5 times more likely to shed cysts than calves in smallholder farms (p=0.037). Calves with diarrhea were nearly three times more likely to be positive for Cryptosporidium oocysts than those with normal feces (p=0.027). At the animal level, larger farms and younger calves were associated with Giardia cysts shedding, while larger herd size and lose fecal consistency were associated with Cryptosporidium oocysts shedding. Giardia and Cryptosporidium infection are endemic in the studied dairy farms. Therefore, detailed molecular epidemiological studies are essential to identify the role of domestic animals in the transmission of infections to humans and vice versa, and to determine the best options for prevention and control of cryptosporidiosis and giardiasis.

Outbreak of Brucella melitensis infection in Eastern Sicily: risk factors, clinical characteristics and complication rate.

Brucellosis is one of the most common zoonoses in the world, especially in Southern Italy, where many cases are still recorded every year. 128 cases of brucellosis were observed in Messina (Sicily) in 2016, representing a tenfold increase in the number of cases of brucellosis expected. The aim of this multicenter retrospective study was to analyze clinical and microbiological aspects of a brucellosis outbreak in the province of Messina in 2016, the incidence of its complications and the treatment combinations applied. The principal transmission route was through the ingestion of unpasteurized fresh cheese. The mean latency period between the onset of the symptoms and diagnosis was 35.58±42.75 days. A late diagnosis increases the risk of developing complications. Drug-resistant strains of B. melitensis to Trimethoprim/ Sulfamethoxazole and Ciprofloxacin were found in blood cultures of 58.4% patients. Brucellosis is still present in Sicily. A diagnostic delay predisposes to complications requiring prolonged therapies. The finding of Brucella melitensis strains resistant to the most widespread treatments is worrisome and needs further investigation. Moreover, the use of alternative combination antibiotic therapy is recommended.

Outbreak of Brucella melitensis infection in Eastern Sicily: risk factors, clinical characteristics and complication rate.

Brucellosis is one of the most common zoonoses in the world, especially in Southern Italy, where many cases are still recorded every year. 128 cases of brucellosis were observed in Messina (Sicily) in 2016, representing a tenfold increase in the number of cases of brucellosis expected. The aim of this multicenter retrospective study was to analyze clinical and microbiological aspects of a brucellosis outbreak in the province of Messina in 2016, the incidence of its complications and the treatment combinations applied. The principal transmission route was through the ingestion of unpasteurized fresh cheese. The mean latency period between the onset of the symptoms and diagnosis was 35.58 ± 42.75 days. A late diagnosis increases the risk of developing complications. Drug-resistant strains of B. melitensis to Trimethoprim/Sulfamethoxazole and Ciprofloxacin were found in blood cultures of 58.4% patients. Brucellosis is still present in Sicily. A diagnostic delay predisposes to complications requiring prolonged therapies. The finding of Brucella melitensis strains resistant to the most widespread treatments is worrisome and needs further investigation. Moreover, the use of alternative combination antibiotic therapy is recommended.

Toxoplasmosis in camels (Camelus dromedarius) of Borana zone, Oromia region, Ethiopia: seroprevalence and risk factors.

Toxoplasmosis is one of the most prevalent parasitic infections of medical and veterinary importance. A cross-sectional study was conducted from November 2013 to January 2014 to estimate the seroprevalence of Toxoplasma gondii infection in camels from four districts of Borana zone, Southern Oromia, Ethiopia. In addition, a questionnaire survey was administered to 124 pastoralists to identify possible risk factors and to assess the awareness level of pastoral communities about toxoplasmosis. A total of 396 serum samples were examined for anti-Toxoplasma IgG antibodies using the direct agglutination test (DAT). Fisher's exact test and logistic regression were used for data analysis. An overall seroprevalence of 8.33 % (95 % confidence interval [CI] 5.60 %, 11.07 %) at animal-level and 37.5 % (95 % CI: 20.1 %, 57.4 %) at herd-level was found. The seroprevalence was significantly high in Moyale district (23.07 %) followed by Yabello (7.20 %), Dirre (3.77 %), and Arero (0.0 %) districts (P < 0.001). Multivariable logistic regression analysis revealed that the likelihood of acquiring T. gondii infection was significantly higher in camels of Moyale district (adjusted OR = 5.89, 95 % CI 2.15, 16.12; P = 0.001) than Dirre district, in camels of >8 years old (adjusted odds ratio [OR] = 4.95, 95 % CI 1.68, 14.55; P = 0.004) than camels of ≤4 years old. There was no significant association between herd-level seroprevalence of T. gondii infection and abortion history, herd size, and presence of domestic cats and wild felids (P > 0.05). The majority of interviewees were uneducated (82.25 %), and all had no knowledge of toxoplasmosis. All camel herders drink raw camel milk but consume cooked meat (90.32 %). Of the interviewees, 93.06 % are aware about soil-eating habit of camels and provide salt supplement for their camels. Majority of the respondents practice improper disposal of aborted materials (throw along the way) (88.70 %), and 73 % of the study participants do not wash their hands after handling aborted fetus. The results of the present study confirm relatively lower prevalence of T. gondii infection in camels reared in Borana zone. Age and study district are significant predictors of T. gondii seropositivity. The vast majorities of interviewed pastoralists were uneducated and practice poor biosecurity measures to prevent diseases. Education of pastoralists about biosecurity measures to prevent toxoplasmosis and further studies are warranted to unravel the economic and public health consequences of T. gondii infection.

First report on seroepidemiology of Toxoplasma gondii infection in pigs in Central Ethiopia.

BACKGROUND: Toxoplasma gondii is one of the most widely prevalent cyst forming Apicomplexan parasites with significant impact on animal production particularly in sheep, goats and pigs. The objectives of this cross-sectional study were to estimate the seroprevalence and to assess risk factors of Toxoplasma gondii infection in pigs. A systematic random sampling technique was used to collect 402 blood samples from pigs in Central Ethiopia. Direct Agglutination Test (DAT) was used to test sera. A questionnaire survey was made to assess potential risk factors and knowledge of farm attendants about toxoplasmosis. RESULTS: An overall seroprevalence of 32.1% [95% confidence interval (CI): 27.6%-36.9%] was found. Multivariable logistic regression analysis showed that extensively managed pigs (39.7%) are nearly twice (adjusted odds ratio [aOR]:=1.91, 95% CI: 1.01, 3.63) at higher risk of acquiring toxoplasmosis than intensively managed pigs (30.5%). Pigs supplied with feed containing animal byproducts had nearly four times (OR = 3.84, 95% CI: 2.01, 7.36) higher risk of acquiring T. gondii infection. Most of the farm attendants had little knowledge of health risks due to cats, neither to human nor to animals. Absence of rodent control, high neonatal mortality and history of abortion were found among herds of the studied pig farms. CONCLUSIONS: T. gondii infections in pigs are wide spread. Extensive management systems and pig feed types containing animal byproducts are independent predictors of T. gondii seropositivity. The high seroprevalence suggests that pigs might serve as an important source of T. gondii infection for people. This is the first report of seroepidemiology of T. gondii infection in pigs in Ethiopia. Further studies are warranted for designing appropriate prevention and control strategies.

First report of Toxoplasma gondii in camels (Camelus dromedarius) in Ethiopia: bioassay and seroepidemiological investigation.

BACKGROUND: Toxoplasmosis is a major public health concern in many countries of the world. A cross-sectional and follow up experimental study designs were used for seroepidemiological and bioassay studies, respectively from November 2012 to April 2013. The objectives were to estimate the seroprevalence of T. gondii infection, to assess risk factors and to isolate the parasite from camels in the Fentale district, Ethiopia. A direct agglutination test (DAT) and indirect enzyme linked immunosorbent assay (ELISA) kits were used to test camel sera. Hearts and tongues (each 25 g) from 31 seropositive camels were bioassayed in mice. Associations between seroprevalence and potential risk factors (collected using a questionnaire survey) were analyzed using logistic regression. RESULTS: An overall T. gondii prevalence of 49.62% (220/455) by DAT and 40.49% (179/451) by indirect ELISA test were detected. Herd level seroprevalence of 96.77% (30/31) (95% CI: 83.30- 99.92) by DAT was recorded and it was significantly higher in areas where wild felids are present (P = 0.038). Multivariable logistic regression showed that the likelihood of acquiring T. gondii infection was significantly higher in camels in the Ilala pastoral association [PA] (82.26%) (Adjusted Odds ratio [aOR] = 10.8; P < 0.001) than camels in the Galcha PA (31.43%), in camels of ≥ 8 years old (56.52%; aOR = 1.88; P = 0,033) than camels of ≤ 4 years old (34.26%) and in areas where domestic cats are present (aOR = 4.16; P = 0.006). All camel owners were uneducated, handle aborted fetus with bare hands, and drink raw camel milk. DAT and ELISA tests had moderate agreement (Kappa = 0.41). Viable T. gondii were isolated from 16.13% (5/31) of DAT positive camels. One DAT positive but ELISA negative camel sample gave a cyst positive result. CONCLUSIONS: T. gondii infection of camels in the study district is widespread. Age, presence of domestic cats and study PA are independent predictors of T. gondii seropositivity. Isolation of viable parasites from edible tissues of camels and the very poor knowledge of pastoralists about toxoplasmosis suggest the need for prevention of toxoplasmosis through bio-security measures, education and further investigation to unravel the impact of camel toxoplasmosis deserves consideration.

Majority of T. gondii seropositive chickens (Gallus domesticus) in Central Ethiopia carries the infective parasite.

BACKGROUND: The prevalence of Toxoplasma gondii in free range chickens is a good indicator of the prevalence of T. gondii oocysts in the environment. The aim of this study was to isolate T. gondii parasites from heart and brain of seropositive free range (FR) chickens. FINDINGS: Isolation of T. gondii from pooled heart and brain of 41 direct agglutination test (DAT) positive (≥1:40) free range chickens (Gallus domesticus) was carried out by bioassay in mice. T. gondii specific antibodies in mice were assayed by DAT and microscopy was employed for detection and enumeration of brain tissue cysts. Overall, bioassay was positive in 29 (70.7%) chicken samples. T. gondii tissue cysts were isolated from 59% (24/41) of bioassayed chickens: from 2 of 7 chickens with a titer of 1: ≤ 60, 2 of 5 with titer 1: 180, 6 of 8 with titer 1: 540, 10 of 15 with titer 1: 1620, 1 of 2 with titer 1: 6000, 2 of 3 with titer 1:18000, 1 of 1 with titer 1:54000. None of the isolates was pathogenic for mice. Tissue cysts were detected from 61% of seropositive mice (DAT ≥ 1:40). Generally, tissue cyst counts per brain of mouse were low (mean: 132.7 ± 84.4; range: 47-352). CONCLUSIONS: Majority of T. gondii seropositive chickens (Gallus domesticus) in Central. Ethiopia carries the infective parasite. Tissues from the free range chicken might be a source infection for animals and humans.

Some risk factors for reproductive failures and contribution of Toxoplasma gondii infection in sheep and goats of Central Ethiopia: a cross-sectional study.

Reproductive failure has a negative impact on animal production, health and welfare and ultimately rural economies. In Ethiopia, the factors affecting small ruminant reproductive efficiency are inadequately investigated. A cross-sectional survey was done from November, 2010 to May, 2011 to investigate risk factors of reproductive failures including Toxoplasma gondii infection. Analysis of 1372 sera (787 sheep and 585 goats) from 409 flocks using ELISA showed high flock (59.7%) and animal (31.8%) level T. gondii seroprevalence. An overall 24.9% (341/1372) annual abortion rate (19.6% in sheep and 32.0% in goats) was recorded. Animal level T. gondii seroprevalence was significantly associated with abortion in Ambo and Ada'a-Liben districts (Odds ratio [OR] = 2.31, 95% confidence interval [CI]: 1.42, 3.76; P = 0.001). Questionnaire survey on 199 households of Ambo, Ada'a-Liben and Fentale districts revealed high flock level abortion (57.5%), still birth (28.9%) and neonatal losses (47.9%), which are significantly different between study districts (P<0.001). Multivariable logistic regression model showed that species (goat), large flock size, pastoral and agro-pastoral production systems, drinking water from river and stagnant water bodies, grazing/browsing in plain land and extensive management were significant predictors of flock level reproductive failures. In the final Zero inflated Poisson regression model number of abortions was significantly higher in goat than in sheep flocks (Incidence risk ratio [IRR] = 1.42, 95% CI: 1.08, 1.87; P = 0.012). It is also significantly higher in pastoral (IRR = 1.81, 95% CI: 1.16, 2.83; P = 0.008) and agro-pastoral production systems (IRR = 1.94, 95% CI: 1.32, 2.86; P = 0.001) than in sedentary production system. Prevention of toxoplasmosis, improved husbandry practices and further epidemiological studies to identify causes of reproductive failures are recommended.

Seroepidemiological study of ovine toxoplasmosis in East and West Shewa Zones of Oromia Regional State, Central Ethiopia.

BACKGROUND: Toxoplasmosis is a globally distributed zoonosis. Consumption of raw or undercooked meat, which is among the main risk factors for acquiring human infection, is a popular tradition in Ethiopia. However, studies on toxoplasmosis in food animals used for human consumption in Ethiopia are very scarce. Thus, the objectives of the present study were to estimate the seroprevalence and the risk factors of T. gondii infection in sheep in Ambo, Ada'a-Liben and Fentale districts of Central Ethiopia. Sera from 1130 sheep were analyzed for Toxoplasma gondii specific IgG antibodies using an indirect enzyme linked immunosorbent assay (ELISA) with the P30 antigen. A questionnaire was administered to assess potential risk factors for T. gondii seropositivity. Association of seroprevalence with potential risk factors related to altitude, host and farm characteristics were analyzed by univariable and multivariable logistic regression. RESULTS: Overall flock and animal level seroprevalences were 70.48% (160/227; 95% CI: 64.51, 76.46) and 31.59% (357/1130; 95% CI: 28.88, 34.31), respectively. The multivariable logistic regression model indicated that the probability of acquiring T. gondii was higher in sheep from highland (2300 - 3200 meters above sea level) [Odds ratio (OR) = 4.11, 95% confidence interval (CI): 2.65, 6.36; P < 0.001] and midland (OR = 4.54, 95% CI: 2.76, 7.49; P < 0.001) than from lowland (<1500 meters above sea level), in females than in males (OR = 1.60, 95% CI: 1.04, 2.43, P = 0.033), in adult than in young animals (OR = 2.93, 95% CI: 1.97, 4.35, P < 0.001), in small than in large flocks (OR = 3.34, 95% CI: 1.26, 8.86, P = 0.016), and in sheep that were given tap water (OR = 4.07, 95% CI: 1.07, 15.42, P = 0.039) and river water (OR = 4.18, 95% CI: 1.54, 11.35, P = 0.005) than in those that drunk water from mixed sources (i.e., river, well, lake and pond). CONCLUSIONS: The high flock and animal level seroprevalence of toxoplasmosis in sheep is a good marker of the potential risk for human infections. Altitude, sex, age, flock size and source of water were identified as important risk factors to acquire the infection. Public education and awareness training are imperative in order to alleviate the danger posed to consumers. Further detailed studies to assess the impact of infections are warranted.

Seroepidemiology of Toxoplasma gondii infection in women of child-bearing age in central Ethiopia.

BACKGROUND: Toxoplasma gondii infections during pregnancy can result in abortion or congenital defects. Prevalence and risk factors of toxoplasmosis in women of child-bearing age in Ethiopia are unknown. The current study was conducted with the objectives of estimating the seroprevalence and potential risk factors in acquiring T. gondii infection by women of child-bearing age in Central Ethiopia. METHODS: A cross-sectional study was conducted from March 2011 to September 2011. Sera of 425 women were analyzed by indirect enzyme linked immunosorbent assay (ELISA). A questionnaire survey was administered for all study participants to gather information on risk factors. RESULTS: The study revealed that anti- T. gondii IgG antibodies were detected in 81.4% of the samples of which 78.4% were positive for only IgG and 3.06% positive for both IgG and IgM antibodies. Seroprevalence of IgM antibodies to T. gondii (4.0%, 95% CI: 2.14, 5.86) was suggestive of recent infections. Of the 213 pregnant women 9 (4.2 %) were IgM reactive. Out of 17 potential risk factors investigated, univariate logistic regression showed significant association of T. gondii infection with study area, age, pregnancy status, raw vegetable consumption, source of water, presence of cats at home, contact with cats, HIV status and precaution during cats' feces cleaning (P ≤ 0.05). The final logistic regression model revealed that: the probability of acquiring T. gondii infection by women of Debre-Zeit was 4.46 times (95% CI of adjusted odds ratio [aOR]: 1.67, 11.89; P =0.003) higher compared to women of Ambo, pregnant women were twice (95% CI aOR: 1.13, 3.59; P = 0.018) more likely to be seropositive than non-pregnant women and women who consume raw vegetable were at increased risk of infection (aOR = 2.21, 95% CI: 1.03, 4.78; P = 0.043) than women who didn't consume. CONCLUSION: The seroprevalence of T. gondii infection in women of child-bearing age in Central Ethiopia is high. Study area, pregnancy and raw vegetable consumption are risk factors to acquire T. gondii infection. Educational program, antenatal screening of pregnant women and further epidemiological studies to uncover the economic and health impact of toxoplasmosis are suggested.

Seroepidemiological study of caprine toxoplasmosis in East and West Shewa Zones, Oromia Regional State, Central Ethiopia.

Toxoplasmosis is a global zoonosis caused by Toxoplasma gondii, an intracellular apicomplexan parasite. The objectives of this study were to estimate the animal and flock level seroprevalence and risk factors associated with toxoplasmosis in goats of Central Ethiopia. In Ethiopia, goats are economically important animals used for meat and milk production. The study was cross-sectional and 927 blood samples from 187 goat flocks were collected to examine T. gondii specific IgG antibodies by enzyme linked immunosorbent assay (ELISA). A questionnaire was used to collect data on the potential risk factors. The study revealed flock and animal level seroprevalence of 58.3% (109/187; 95% confidence interval [CI]: 51.16, 65.42) and 19.7% (183/927; 95% CI: 17.17, 22.31), respectively. The likelihood of acquiring T. gondii infection was higher in semi-intensively managed goats (Odds ratio [OR]=2.48, 95% confidence interval [CI]: 1.14, 5.37; P=0.022) than in extensively managed goats, in females than in males (OR=1.84, 95% CI: 1.16, 2.92; P=0.010), in adults than in young animals (OR=2.00, 95% CI: 1.21, 3.28; P=0.006), in small than in large flocks (OR=2.03, 95% CI: 1.03, 3.98; P=0.040), in goats kept under sedentary (OR=2.81, 95% CI: 1.41, 5.59; P=0.003) and agropastoral farming system (OR=3.62, 95% CI: 1.83, 7.18; P<0.001) than pastoral farming system and in goats allowed to drink water from the tap than those drinking from river and stagnant water bodies (OR=9.25, 95% CI: 3.04, 28.15; P<0.001). Our study indicates that exposure of goats to oocysts of T. gondii is widespread. We recommend further studies to determine the genotype of the parasite, public health and economic impacts of toxoplasmosis and the role of raw goat meat and milk as a source of infection for consumers.

Development and validation of two PCR tests for the detection of and differentiation between Anaplasma ovis and Anaplasma marginale.

Anaplasma ovis and Anaplasma marginale are tick-transmitted bacteria that cause anaplasmosis in domestic and wild animals. Recent results show that some domestic and wild animals and ticks are susceptible to both A. ovis and A. marginale, thus supporting the need to differentiate between these species in hosts and ticks diagnosed with Anaplasma infection. However, although anaplasmosis is one of the most common diseases of grazing animals worldwide, rapid and effective tests are not available for the detection of and discrimination between these 2 Anaplasma species. The objective of this research was to develop an easy and reliable method to identify and discriminate between the closely related pathogens A. ovis and A. marginale. A. ovis and A. marginale major surface protein 4 (msp4) gene sequences were retrieved from different geographic strains and aligned to design 2 sets of primers in a region with significant differences between the 2 species, but completely conserved among strains. PCR reactions using these primers were 100% species-specific and detected all strains from each pathogen previously identified with other methods. The 2 sets of primers designed for the specific PCR amplification of A. ovis and A. marginale allow easy-to-detect and discriminate between the 2 pathogens, thus avoiding the time-consuming sequencing or multi-gene amplification procedures. This PCR provides a tool for the detection of A. ovis and A. marginale in ticks and in wildlife and domestic hosts.

Epidemiological significance of the domestic black pig (Sus scrofa) in maintenance of bovine tuberculosis in Sicily.

Bovine tuberculosis (bTB) is an emerging disease among wild animals in many parts of the world. Wildlife reservoir hosts may thus represent a potential source of infection for livestock and humans. We investigated the role played by the Sicilian black pig, an autochthonous free- or semi-free-ranging domestic pig breed, as a potential source of bTB infection in an area where bTB prevalence in cattle is high. We initially performed a preliminary field study to assess the occurrence of bTB in such animals. We sampled 119 pigs at abattoir and found 6.7% and 3.4% of them to be affected by gross tuberculous-like lesions (TBL) and Mycobacterium bovis culture positive, respectively. We then proceeded to investigate the dissemination and characteristics of lesions in a second field study performed on 100 animals sampled from infected herds. Here, tissues collected at the abattoir were examined macroscopically, microscopically, and by culture tests. Most pigs with TBL showed generalized lesions in both gross and histological examinations (53% and 65.5%, respectively). Head lymph nodes were the most frequently affected in both localized and generalized TB cases observed macroscopically and microscopically. M. bovis was the most frequently isolated etiologic agent. The molecular characterization of isolates from both field studies by spoligotyping and analysis of 12 mycobacterial interspersed repetitive-unit-variable number tandem repeat (MIRU-VNTR) loci, followed by their comparison to isolates of cattle origin, suggested a potential transmission of mycobacteria from domestic animals to black pigs and vice versa. Our findings, along with ethological, ecological, and management considerations, suggest that the black pig might act as a bTB reservoir in the ecosystem under study. However, additional studies will be necessary to establish the true epidemiological significance of the Sicilian black pig.

Bioaccumulation of dioxin-like substances and selected brominated flame retardant congeners in the fat and livers of black pigs farmed within the Nebrodi Regional Park of Sicily.

An observational study was designed to assess the bioaccumulation of polychlorodibenzodioxins (PCDD) and polychlorodibenzofurans (PCDF), dioxin-like polychlorobiphenyls (DL-PCB), and 13 selected polybromodiphenylethers (PBDE) in autochthonous pigs reared in the Nebrodi Park of Sicily (Italy). Perirenal fat and liver samples were drawn from animals representative of three different outdoor farming systems and from wild pigs and then analyzed for the chemicals mentioned previously. The highest concentrations of PCDD + PCDF and DL-PCB were detected in the fat (0.45 and 0.35 pg World Health Organization toxicity equivalents [WHO-TE] per g of fat base [FB], respectively) and livers (12.7 and 3.28 pg WHO-TE per g FB) of the wild group, whereas the free-ranging group showed the lowest levels (0.05 and 0.03 pg WHO-TE per g FB in fat and 0.78 and 0.27 pg WHO-TE per g FB in livers). The sum of PBDE congeners was highest in wild pigs (0.52 ng/g FB in fat and 5.64 ng/g FB in livers) and lowest in the farmed group (0.14 ng/g FB in fat and 0.28 ng/g FB in livers). The contamination levels in fat and livers of outdoor pigs had mean concentration values lower than those levels reported for intensively indoor-farmed animals. In wild pigs, bioaccumulation was associated with their free grazing in areas characterized by bush fires. The results of this study aid to emphasize the quality of the environment as a factor to guarantee food safety in typical processed pig meat products, specifically from outdoor and extensive Nebrodi farming systems.

A case of generalized bovine tuberculosis in a sheep.

The present report describes a rare case of generalized bovine-type tuberculosis in a slaughtered 4-year-old ewe discovered during routine surveillance at an abattoir. A postmortem examination revealed lesions in the ewe's thoracic and abdominal cavities, ranging from encapsulated, mineralized foci to extensive, soft, caseous tissue. Lesions in the lungs, liver, and lymph nodes were consistent with mycobacterial infection. A histopathological examination detected granulomatous lesions in all tissue samples. The presence of Mycobacterium tuberculosis complex genome was confirmed through a polymerase chain reaction (PCR) analysis of tissues, using IS6110 primers, followed by a nucleotide sequence analysis of PCR products. Acid-fast bacteria, characterized as Mycobacterium bovis, were isolated from lesions following 38 days of incubation.

Characterization of Anaplasma phagocytophilum and A. ovis infection in a naturally infected sheep flock with poor health condition.

Anaplasma species are transmitted by ticks and cause diseases in humans and animals. These pathogens infect sheep, an economically important domestic animal worldwide. The current study was designed to characterize in 200 animals the infection with Anaplasma phagocytophilum and Anaplasma ovis and the genetic diversity of A. ovis strains collected from a naturally infected sheep flock with poor health condition. Sheep had 98% seroprevalence to Anaplasma spp. antibodies. PCR results confirmed the presence of A. phagocytophilum and A. ovis DNA in 11.5% and 37% of the sheep, respectively. Concurrent infections were detected in 6.5% of the sheep. Seventy-one adult ticks were collected from 45 sheep with infestations ranging from one to 15 ticks per animal. The analysis of A. ovis msp4 sequences demonstrated a previously unreported polymorphism for this pathogen with 17 different haplotypes in infected sheep. These results demonstrated that, although A. ovis msp4 haplotypes may be less variable when compared with Anaplasma marginale and A. phagocytophilum strains on a global scale, genetic polymorphisms occur in this locus in strains obtained from an infected sheep flock with poor health condition.

Natural trematode infestation in feral Nebrodi Black pigs: pathological investigations.

Few studies describe the parasites of pigs bred in the wild state, although pigs are a known reservoir of trematode infestation. This article reports the results of a retrospective study carried out from January 2003 to June 2007 on 3021 Nebrodi Black male and female pigs, regularly slaughtered, aged between 8 months and 4 years. Fasciola hepatica and Dicrocoelium dendriticum flukes were detected in 143 (4.37%) of 3021 livers. The predominant histological features were multifocal to diffuse chronic hepatitis, with fibrosis and severely thickened walls of the bile ducts and chronic parietal, sometimes nodular inflammation. F. hepatica infestation was frequently associated with marked hyperplasia and hypertrophy of the submucosal glands. The study results confirm the important role swine play in the transmission of trematode infestations, indicate the prevalence of these parasites in the Nebrodi Park area, and draw attention to the need for a prophylaxis plan to prevent the spread of infestation to ruminants and humans living in the area.

Molecular epidemiological and antibiotic susceptibility characterization of Brucella isolates from humans in Sicily, Italy.

Brucellosis is a serious problem in Sicily. Brucella melitensis was identified as the species most frequently isolated in humans in Italy. No data, however, are available about the molecular epidemiological characterization of Brucella isolates from humans. We have conducted this study to molecularly characterize clinical isolates of Brucella spp. and to evaluate their antimicrobial susceptibilities. Twenty Brucella isolates were studied. Differential growth characteristics and DNA polymorphisms such as the restriction patterns of the PCR-amplified omp2a and omp2b genes, rpoB nucleotide sequencing, and multiple-locus variable-number tandem repeat analysis of 16 loci (MLVA-16) were used to characterize the strains. In vitro antibiotic susceptibility was determined by the E-test method on two different agar media, and the results were compared. All isolates were identified as B. melitensis biovar 3. rpoB nucleotide sequence analysis allowed the identification of two different genotypes of B. melitensis biovar 3. On the other hand, the MLVA-16 typing assay recognized 17 distinct genotypes. All isolates were sensitive to all tested antibiotics (rifampin, doxycycline, ciprofloxacin, ceftriaxone, and trimethoprim-sulfamethoxazole), and the Mueller-Hinton agar plate is recommended for antibiotic susceptibility testing by the E-test method. Our findings identify B. melitensis biovar 3 as the etiological agent isolated in Sicily and encourage the use of both molecular methods, and in particular of the MLVA-16 assay, in epidemiological trace-back analysis. This study represents the first epidemiological data from molecular typing of Brucella strains circulating in Italy and, in particular, in eastern Sicily.

Polymerase chain reaction method for leptospirosis, analysis on samples from an autochthon swine population in Sicily, Italy.

We set a method targeting 16 rRNA gene consisting in a single polymerase chain reaction of 40 cycles which is specific for pathogenic leptospira. Negative polymerase chain reaction results were observed with nonpathogenic Leptospira (serovar patoc) and other bacteria species. By this method a survey on a population of autochthon swine herds had been conducted in Sicily particularly on kidney samples of slaughtered animals and on urine samples from live animals. The analysis showed that a prevalence of leptospira up to 40 % can be observed on these animals. Results on other bovine and ovine herds from the same province in Sicily showed a lower prevalence.