Bovine Besnoitiosis Research Articles

Blurred epidemiology of bovine besnoitiosis: parasite DNA detection in skin among seropositive cattle

Blurred epidemiology of bovine besnoitiosis: parasite DNA detection in skin among seropositive cattle

Epidemiologic study on Besnoitia besnoiti infection in dairy herds in Jordan.

Besnoitia besnoiti is an apicomplexan parasite and the causative agent of bovine besnoitiosis which is considered as a re-emergent disease in Europe. A cross-sectional serological study was conducted to determine the seroprevalence and to identify risk factors associated with B. besnoiti infection in 68 dairy herds (n = 806 cows) in Jordan during the period from January to June 2007 and the spring of 2014. Data regarding herd's management was obtained by filling questionnaires through personal interviews with farmers. An indirect ELISA test was used to detect antibodies against B. besnoiti. Chi-square analysis and multivariable logistic regression model were used to identify risk factors associated with seropositivity to B. besnoiti. At the individual cow and herd level, the true prevalence of seropositive animals was 6 and 28.7%, respectively. Cows between 2 and 6years of age had significantly higher seroprevalence of B. besnoiti than other age groups. The highest seroprevalence of B. besnoiti was found in Zarqa and Irbid governorates. Multivariable logistic regression model identified that exchanging visits by farm workers to neighboring farms as a risk factor for seropositivity to B. besnoiti, while smaller herd size and twice a day farm cleaning using sweeping and water hosing were identified as protective factors. This is the first study that investigated the seroprevalence of B. besnoiti infection in dairy herds in Jordan. Further studies are warranted to explore the clinical manifestation of B. besnoiti infection as well as to identify the possible presence of other Besnoitia species and definitive hosts for the parasite.

Besnoitia besnoiti infection in cattle and mice: ultrastructural pathology in acute and chronic besnoitiosis

Current knowledge on bovine besnoitiosis, caused by the emerging apicomplexan pathogen Besnoitia besnoiti, is still fragmentary. So far, studies dealing with ultrastructural pathology focused mainly on the easily accessible chronic stage, whereas ultrastructural investigations of tachyzoites were confined to in vitro studies. In the study presented here, the ultrastructural pathology of naturally B. besnoiti-infected cattle in the acute and chronic disease stages and experimentally B. besnoiti-infected mice was monitored. Further, the ultrastructure of tachyzoites and bradyzoites was investigated. Skin samples of two adult Limousin cows and one adult Limousin bull naturally infected with B. besnoiti and liver and skin samples of gamma-interferon knockout mice infected with B. besnoiti were examined in semithin sections stained with toluidine blue and safranin and in ultrathin sections contrasted with uranyl acetate and lead citrate. Samples of vessel walls of the bull and nasal mucosa of one cow were examined by scanning electron microscopy. Few tachyzoites-like endozoites were detected for the first time in bovine skin, and large numbers of tachyzoites were detected in murine skin and liver. Within tissue cysts in bovine skin, numerous bradyzoites were observed displaying signs of degeneration. Tachyzoites had apicomplexan endozoite ultrastructure. B. besnoiti tachyzoites and bradyzoites differed in shape and the number of amylopectin granules. Transmission and scanning electron microscopy confirmed the presence of two different cyst wall layers, and the present results on cyst wall ultrastructure were in accordance with those previously obtained by histological sections.

Natural Besnoitia besnoiti infections in cattle: hematological alterations and changes in serum chemistry and enzyme activities

BackgroundThe emerging disease bovine besnoitiosis is caused by the apicomplexan parasite Besnoitia besnoiti. Clinical signs of acute besnoitiosis are pyrexia, anorexia and subcutaneous edema. In subacute and chronic besnoitiosis parasitic cysts arise in a variety of tissues and affected cattle display skin lesions and weight loss. In all stages of bovine besnoitiosis, lesions can be found in many organ systems and therefore presumably alter a variety of laboratory parameters. In this study, the impact of naturally acquired acute, subacute and chronic bovine besnoitiosis on hematologic parameters, serum chemistry, and enzyme activities was investigated. Laboratory parameters of two Simmental heifers and two Limousin cows were monitored during acute, subacute and chronic besnoitiosis and in another Simmental heifer during subclinical besnoitiosis. To determine aberrations of laboratory parameters, values were compared with reference ranges obtained from B. besnoiti negative Simmentals (224 samples of nine animals) and Limousins (41 animals). Further, laboratory parameters of B. besnoiti seropositive Limousin cows (54 animals; 32 of these showing clinical signs) and healthy B. besnoiti seronegative Limousin cows (41 animals) were compared.ResultsDuring acute and subacute besnoitiosis, a reduction of leukocyte and erythrocyte concentrations, hematocrit, serum albumin, urea, magnesium, and calcium concentrations were observed. Serum total protein, globulin, total bilirubin and creatinine concentrations were increased and aspartate transaminase (AST) and creatine kinase (CK) activities were elevated. In chronic besnoitiosis, erythrocyte parameters were statistically significantly lower, and total protein and globulin concentrations were significantly higher in B. besnoiti seropositive compared with B. besnoiti seronegative Limousin cows.ConclusionsIn this study, altered laboratory parameters during the course of naturally acquired acute, subacute and chronic bovine besnoitiosis are described for the first time. Only a few animals were examined in acute and subacute besnoitiosis, however the alterations of laboratory parameters during these stages reflected i) the acute inflammatory state (e.g. high levels of serum globulin fractions), ii) clinical findings such as disturbed condition (e.g. bilirubin concentrations), and iii) lesions such as muscle necroses described in the literature (e.g. AST or CK activities). Chronic besnoitiosis led to typical alterations of chronic inflammatory diseases like hyper-(gamma)-globulinemia or reduced erythrocyte concentrations.

Natural Besnoitia besnoiti infections in cattle: chronology of disease progression.

BackgroundBovine besnoitiosis is an emerging protozoan disease in cattle. Neither vaccines nor chemotherapeutic drugs are currently available for prevention and treatment of Besnoitia besnoiti infections. Therefore the implementation of appropriate disease management strategies is of utmost importance. The aim of this longitudinal study was to complement current knowledge on the chronology of disease progression. This was realized by correlating clinical findings in early stages of naturally acquired bovine besnoitiosis with results of real-time PCR of skin biopsies and of two western immunoblots and an immunofluorescent antibody test (IFAT).Animals for this study were obtained by i) closely monitoring a cow-calf operation with a high prevalence of bovine besnoitiosis for cases of acute disease, and by ii) conducting a 12-week cohabitation experiment on pasture with five healthy heifers, a healthy bull and five B. besnoiti infected cows. A control group of six healthy heifers was kept at a minimal distance of 20 m. Further, the spectrum of potential insect vectors was determined.ResultsInfected cattle were followed up to a maximum of 221 days after first detection of B. besnoiti antibodies. Two severely affected cows developed visible and palpable alterations of skin, a decrease in body condition despite good feed intake, and chronic bovine besnoitiosis-associated laminitis leading to non-healing sole ulcers. The cows also had high reciprocal IFAT titers and high loads of parasite DNA in skin samples. Two heifers developed a mild clinical course characterized by few parasitic cysts visible in the scleral conjunctivae and vestibula vaginae. Both heifers became infected during the time of high insect activity of the species Musca domestica, Musca autumnalis, Haematobia irritans, and Stomoxys calcitrans. When a third heifer became subclinically infected, low insect activity was recorded. None of the six control heifers contracted a B. besnoiti infection.ConclusionsIn chronic besnoitiosis, the severe clinical course apparently corresponded with high reciprocal IFAT titers and high loads of parasite DNA in skin, whereas mild and subclinical cases displayed lower values. Bovine besnoitiosis-associated laminitis represents an important complication in severe chronic disease which severely impairs animal welfare.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-015-0344-6) contains supplementary material, which is available to authorized users.

Besnoitia besnoiti among cattle in insular and northwestern Italy: endemic infection or isolated outbreaks?

BackgroundBovine besnoitiosis, caused by the apicomplexan Besnoitia besnoiti, is a chronic and debilitating disease considered as emerging in Europe. In Spain, Portugal and France it is endemic and foci of infection were recorded in Germany, Switzerland, Hungary, Greece and Italy. In Italy, cases of bovine besnoitiosis were registered both in imported and autochthonous cattle, and mostly in central regions; high seroprevalence was also revealed by an epidemiological survey performed in the southern part of the country. Aiming to update information on the disease in northwestern and insular areas of Italy, where data on bovine besnoitiosis were missing, a serosurvey was designed for the present study.MethodsThree thousand one hundred and forty bovine blood samples from both dairy and beef farms (n = 126) were collected in northwestern regions (Lombardy, Piedmont and Liguria) and in the island of Sardinia. Samples were analyzed by a standardized in-house ELISA and those resulted positive were re-tested by Western Blot (WB) for confirmation. On results obtained by both ELISA and WB, apparent (AP) and true prevalence (TP) were calculated at individual and herd levels. Further, a panel of sera resulted positive to ELISA was analyzed by IFAT.ResultsA total of 712 animals (AP = 22.7%; TP = 18.8%) and 109 farms (AP = 86.5%; TP = 88.2%) showed a positive reaction in ELISA. Only ten (AP = 0.3%; TP = 0%) specimens proceeding from five farms (AP = 3.9%; TP = 1.7%) from Lombardy were confirmed positive to the WB, corresponding to two Holstein Friesian cows and eight beef cattle. IFAT showed a low sensitivity (44.4%) scoring positive in only four samples out of 9 positive to WB.ConclusionsThe survey demonstrated that bovine besnoitiosis cannot still be considered endemic in whole Italy. In fact, independent foci of infection were registered only in Lombardy region. Therefore, a sanitary strategy aimed to increase control measures and to organize monitoring plans, by adequate diagnostic tools is necessary to avoid overestimation of B. besnoiti in Italy.

Naturally acquired bovine besnoitiosis: histological and immunohistochemical findings in acute, subacute, and chronic disease.

The pathogenesis of bovine besnoitiosis, a disease of increasing concern within Europe, is still incompletely understood. In this study, disease progression after natural infection with the causative apicomplexan Besnoitia besnoiti was monitored in histological skin sections of 5 individual female cattle over time. High-frequency skin sampling of 2 cattle with mild and 2 with severe acute, subacute, and chronic besnoitiosis, as well as from 1 animal during subclinical disease, enabled documentation from the beginning of the disease. Skin sections were stained with hematoxylin and eosin and Giemsa, periodic acid-Schiff reaction, and anti-Besnoitia immunohistochemistry. In all 4 clinically affected animals, tachyzoite-like endozoites could be detected for the first time by immunohistochemistry, and tissue cyst evolution was monitored. Besnoitiosis-associated lesions were not detected in the animal showing the subclinical course. Because of the inconsistency of the nomenclature of Besnoitia tissue cyst layers in the literature, a new nomenclature for B. besnoiti cyst wall layers is proposed: tissue cysts consist of a hypertrophied host cell with enlarged nuclei, an intracytoplasmic parasitophorous vacuole with bradyzoites, a sometimes vacuolated inner cyst wall, and an outer cyst wall in more developed cysts. Inner and outer cyst walls can be readily distinguished by using special stains. In 1 animal, extracystic B. besnoiti zoites were immunohistochemically detected during the chronic stage. At necropsy, the 2 severely affected cows displayed large numbers of B. besnoiti cysts in a variety of tissues, including the corium of the claws, contributing mainly to the development of chronic laminitis in these 2 cases.

Prevalence and geographic distribution of Besnoitia besnoiti infection in cattle herds in Portugal.

Bovine besnoitiosis, caused by the apicomplexan parasite Besnoitia besnoiti is considered an emergent disease in Europe. This study aimed to determine the prevalence and geographic distribution of B. besnoiti in cattle herds in continental Portugal and to identify potential spatial clustering of infection. A stratified two-stage cross-sectional serological survey was carried out between March 2012 and May 2013 with the five administrative NUTS II regions, Norte, Centro, Lisboa, Alentejo, and Algarve, as the stratification level. Sera from 391 herds in 220 parishes and 83 municipalities were analyzed by a serial testing strategy, with the modified agglutination test (B-MAT) as the first screening assay and the immunofluorescent antibody test (IFAT) as the confirmatory test. Within-herd prevalence of positive herds varied between 0.7 and 72.4% and was ≥10.3% in half of the infected herds. Using a Bayesian approach, the true prevalence of B. besnoiti in cattle herds was determined to be 5.1% (confidence interval (CI), 3.1-7.8%) and the mean within-herd prevalence of positive herds was 33.0% (CI, 20.3-46.0%). The sensitivity and specificity of the B-MAT were estimated to be 96.9% (CI, 93.7-98.8 %) and 99.7% (CI, 99.6-99.8%), whereas those of the IFAT were 89.6% (CI, 86.0-92.5%) and 99.7% (CI, 98.5-99.9%), respectively. Spatial scan statistics analysis identified one spatial cluster covering the majority of the Alentejo region. Seropositive herds were detected for the first time outside Alentejo, in the region Centro and in the northeast of Portugal. Further epidemiological research is needed to identify eco-biological factors, which could explain the geographic clustering of B. besnoiti in Portugal.

No detection of Besnoitia besnoiti DNA in the semen of chronically infected bulls.

Bovine besnoitiosis is a chronic and debilitating disease observed in many European countries that may cause important economic losses in cattle. The recent widespread of the parasite in Europe had led the European Food Safety Authority to declare bovine besnoitiosis as a re-emerging disease in Europe. Many aspects of the epidemiology of bovine besnoitiosis such as the main routes of transmission are still unclear and need to be further studied. Among the different hypotheses, a sexual transmission has not yet been investigated. Therefore, the aim of this study was to evaluate the presence of Besnoitia besnoiti DNA in the semen of naturally infected bulls by using a highly sensitive method (real-time qPCR). Both pre-sperm and sperm fractions of 40 bulls, including seronegative (n = 11), seropositive subclinically (n = 17), and seropositive clinically (n = 12) infected animals, were collected by electroejaculation and analyzed by real-time qPCR. No B. besnoiti DNA was detected in 27 pre-sperm and 28 sperm fractions of the 40 examined bulls, suggesting that the transmission of B. besnoiti infection by the semen of chronically infected bulls is very unlikely.

Prevalence of <i>Besnoitia besnoiti</i> antibodies in bovine sera and milk in Northern Nigeria

Besnoitia besnoiti , a re-emergent parasite of cattle in Europe, occurs in many countries of Africa and other parts of the world. Clinical observations and incidental findings of B. besnoiti in cattle have been reported in the Southern and Northern regions of Nigeria, but the prevalence of antibodies against this parasite is not yet known. This investigation was designed to determine the seroprevalence of bovine besnoitiosis in Northern Nigeria. A total of 400 cattle were selected at random through cluster sampling of herds from two Local Government Areas (LGA) each, of 5 States in the region (Kano, Kaduna, Katsina, Sokoto and Borno States), between May, 2008 and November, 2009. Sera samples obtained from cows, bulls and calves, and milk from lactating cows with suckling calves were screened with indirect immunofluorescent antibody technique (IFAT) for antibodies to B. besnoiti. Out of the 400 samples 321 (80.3%) were positive for antibodies to B. besnoiti . Cattle sampled in Borno had the highest (87.5%) prevalence of antibodies to B. besnoiti, while those sampled from Katsina State had the least prevalence (62.3%). Wamako LGA of Sokoto State had the highest prevalence of the antibodies (100.0%), while Dan Musa LGA in Katsina had the least prevalence (53.0%) among the ten LGA sampled, however, these differences were not statistically significant (p > 0.05). Similarly, the overall prevalence of antibodies to B. besnoiti did not vary significantly between bulls (84.0%) and cows (79.0%), or in the dry (83.6%) and wet (77.1%) seasons (p > 0.05). The high prevalence of antibodies to B. besnoiti in cattle in Northern Nigeria indicates an endemic state of the disease in this region. Keywords : Antibodies, Besnoitia , Cattle, Northern Nigeria, Prevalence

A review on bovine besnoitiosis: a disease with economic impact in herd health management, caused byBesnoitia besnoiti(Franco and Borges, )

Bovine besnoitiosis is caused by the largely unexplored apicomplexan parasite Besnoitia besnoiti. In cows, infection during pregnancy often results in abortion, and chronically infected bulls become infertile. Similar to other apicomplexans B. besnoiti has acquired a largely intracellular lifestyle, but its complete life cycle is still unknown, modes of transmission have not been entirely resolved and the definitive host has not been identified. Outbreaks of bovine besnoitiosis in cattle were described in the 1990s in Portugal and Spain, and later several cases were also detected in France. More cases have been reported recently in hitherto unaffected countries, including Italy, Germany, Switzerland, Hungary and Croatia. To date, there is still no effective pharmaceutical compound available for the treatment of besnoitiosis in cattle, and progress in the identification of novel targets for intervention through pharmacological or immunological means is hampered by the lack of molecular data on the genomic and transcriptomic level. In addition, the lack of an appropriate small animal laboratory model, and wide gaps in our knowledge on the host-parasite interplay during the life cycle of this parasite, renders vaccine and drug development a cost- and labour-intensive undertaking.

Biting behaviour of Tabanidae on cattle in mountainous summer pastures, Pyrenees, France, and effects of weather variables

In France, during the summer, cattle in mountainous pastures can be highly exposed to tabanid bites. The persistent biting behaviour of tabanids not only causes disturbance, but is also responsible for transmitting diseases, such as bovine besnoitiosis. The purpose of this study was to better identify the level of tabanid annoyance on cattle by means of insect trapping and direct observation of cows. Tabanids were active during the entire daily observation period (10:00-16:00), except for Haematopota sp., which were less active in the morning. The tabanids collected in Nzi traps were generally representative of those that landed on cattle, except for Haematopota sp., as Nzi traps were not very effective for these species. The preferred feeding sites for most species appeared to be cow's legs or udder. Leg stamping was the defensive reaction most related to a tabanid alighting on a host. Generalized linear mixed models showed that the parameters associated with tabanid landings on hosts were related to weather and altitude, but not to landscape structure. Increased landings were mostly associated with the higher temperatures and lower wind speeds at midday, but some differences were observed between species. The results indicate that cattle-protection measures should be taken during the peak of tabanid abundance when climatic conditions favour intense biting activity. Nzi traps set close to livestock were very effective to catch tabanids and could help in reducing the annoyance caused by horse flies.

Seroprevalence of Besnoitia besnoiti infection and associated risk factors in cattle from an endemic region in Europe

Bovine besnoitiosis, caused by the parasite Besnoitia besnoiti, is a chronic, debilitating disease with both cutaneous and systemic clinical signs that has re-emerged in Europe. This is the first random cross-sectional prevalence study of B. besnoiti infection in cattle carried out in an endemic area in Europe (Navarra, Spain). Dairy (n = 372) and beef (n = 340) cattle >1 year of age were randomly blood sampled. Serum was evaluated using a validated ELISA. True animal prevalence data were restricted to beef cattle (16.0%). The prevalence significantly increased with age and seropositive animals were mostly located in mountainous areas where the disease is endemic. Breed and sex were not found to be risk factors.

Prevalence of Besnoitia besnoiti infection in beef cattle from the Spanish Pyrenees

Reports of recent outbreaks of bovine besnoitiosis in Europe have emphasized the need for prevalence studies to determine the importance of the disease. A previous cross-sectional study carried out in a northern province of Spain showed that seropositive cattle were beef breeds primarily located in regions where the disease is endemic (such as in the Pyrenees). The present study focused on this population. Sixty-three beef herds were selected and at least 50% of the animals in each herd were sampled (n = 3318). The herd, intra-herd and animal prevalence rates were calculated. All breeding bulls located in the same region were also sampled (n = 587), and the animal prevalence calculated. Sera were tested by ELISA.The herd prevalence rate of bovine besnoitiosis was 87.3%, and both sexes were similarly affected (approximately 50%). The results indicate that the disease is highly widespread in the Pyrenees, and that serological examination of cattle should be recommended when trading beef cattle.

Besnoitia besnoiti among cattle in insular and northwestern Italy: endemic infection or isolated outbreaks?

Bovine besnoitiosis, caused by the apicomplexan Besnoitia besnoiti, is a chronic and debilitating disease considered as emerging in Europe. In Spain, Portugal and France it is endemic and foci of infection were recorded in Germany, Switzerland, Hungary, Greece and Italy. In Italy, cases of bovine besnoitiosis were registered both in imported and autochthonous cattle, and mostly in central regions; high seroprevalence was also revealed by an epidemiological survey performed in the southern part of the country. Aiming to update information on the disease in northwestern and insular areas of Italy, where data on bovine besnoitiosis were missing, a serosurvey was designed for the present study. Three thousand one hundred and forty bovine blood samples from both dairy and beef farms (n = 126) were collected in northwestern regions (Lombardy, Piedmont and Liguria) and in the island of Sardinia. Samples were analyzed by a standardized in-house ELISA and those resulted positive were re-tested by Western Blot (WB) for confirmation. On results obtained by both ELISA and WB, apparent (AP) and true prevalence (TP) were calculated at individual and herd levels. Further, a panel of sera resulted positive to ELISA was analyzed by IFAT. A total of 712 animals (AP = 22.7%; TP = 18.8%) and 109 farms (AP = 86.5%; TP = 88.2%) showed a positive reaction in ELISA. Only ten (AP = 0.3%; TP = 0%) specimens proceeding from five farms (AP = 3.9%; TP = 1.7%) from Lombardy were confirmed positive to the WB, corresponding to two Holstein Friesian cows and eight beef cattle. IFAT showed a low sensitivity (44.4%) scoring positive in only four samples out of 9 positive to WB. The survey demonstrated that bovine besnoitiosis cannot still be considered endemic in whole Italy. In fact, independent foci of infection were registered only in Lombardy region. Therefore, a sanitary strategy aimed to increase control measures and to organize monitoring plans, by adequate diagnostic tools is necessary to avoid overestimation of B. besnoiti in Italy.

Epidemiological investigations of bovine besnoitiosis in Hungary

It usually gains proper attention, when blood-sucking arthropod species in Europe show northward spreading, in part due to climate change and global warming. Accordingly, it is well known, that the geographical regions where pathogens transmitted by these as biological vectors are endemic, also may extend towards the north. However, it is mostly less emphasized, that for vector-borne pathogens present in Western Europe the risk of eastward emergence is also possible, if they have competent vectors in the central and/or eastern part of our continent. Besnoitia besnoiti (Apicomplexa: Sarcocystidae) is a cyst-forming coccidian parasite that may cause severe lesions (with usually high seroprevalence, but low morbidity and mortality) in cattle as intermediate hosts. Wild ruminants are also susceptible. As a unique example among cystogenic coccidia, the main transmission route of B. besnoiti appears to be mechanical by blood-sucking dipterans (tabanid and stable flies), although it is also possible iatrogenically (with hypodermic needles) and most likely with close contact between animals. Bovine besnoitiosis has been endemic to South-Western Europe for more than a century, but a significant geographical expansion was observed during the last decade to other parts of the formerly endemic countries and to countries neighboring France. During the autumn of 2013 bovine besnoitiosis was diagnosed in a beef cattle herd in Hungary (for the first time in Central-Eastern Europe), following the import of Aubrac heifers and bulls from France in the previous two years. The preliminary serological herd screening with ELISA shows that even after the 2nd year of the presence of imported animals (with high prevalence of B. besnoiti infection), seropositivity among local animals, which originally belonged to the herd, is low. Based on ELISA results venereal transmission (from imported, infected bulls to local, uninfected cows) appears to be either rare or unlikely. The seroprevalence of besnoitiosis decreased significantly among calves born to the group of imported mother cows. The risk of infection seems to be high, when calves stay with their mother during suckling (for 6-7 months), and if animals are kept in the same stable (although physically separated) during the main fly season. Confirmation of the ELISA results is done with immunoblot and IFAT. All seropositive cattle are now kept at a distance of several kilometres from other groups of animals, prior to culling. Molecular and sero-epidemiological evaluation of the situation continues with the aim of preventing the spread of the disease and regaining the epidemiological status of Hungary as exempt of bovine besnoitiosis.

Bovine besnoitiosis emerging in Central-Eastern Europe, Hungary

BackgroundBesnoitia besnoiti, the cause of bovine besnoitiosis, is a cyst-forming coccidian parasite that has recently been shown to be spreading in several Western and Southern European countries.FindingsClinical cases of bovine besnoitiosis were confirmed for the first time in Hungary, by histological, serological and PCR analyses.ConclusionsThis is the first report of autochthonous bovine besnoitiosis in Central-Eastern Europe. The emergence of bovine besnoitiosis in this region represents a further example, when human activity (i.e. cattle trading) is the main factor involved in the geographical spread of an infectious disease.

Evidence for bovine besnoitiosis in Egypt—first serosurvey of Besnoitia besnoiti in cattle and water buffalo (Bubalus bubalis) in Egypt

The present study evaluated the presence of specific antibodies against Besnoitia besnoiti in cattle and water buffalo (Bubalus bubalis) in Egypt. Sera from cattle (n = 216) and water buffaloes (n = 133) collected from five different provinces of Egypt (Behera, Alexandria, Assuit, Gharbia, and Matrouh) were analyzed. Testing for B. besnoiti antibodies by PrioCHECK® Besnoitia Ab 2.0 ELISA initially identified 13.75% (48 out of 349) of individual sera as positive at the manufacturer's suggested cutoff threshold, 15 percent positivity (PP). Statistically significant associations between B. besnoiti prevalence, species, sex, age, and geographical distribution were observed. Seropositive animals were distributed in all of the provinces from which animals were sampled except Gharbia province. Assuit province showed the highest percentage of infection (30.76%) followed by Matrouh, Alexandria, and Behera provinces (25, 16.29, and 9.6%, respectively). The highest infection rate of B. besnoiti was significantly higher in cattle (17.13%) than in water buffaloes (9.02%). Positive cases were observed in all age categories. While the highest infection rate (17.13%) was recorded in the age group 5-10years followed by the age group 1-5years (15.38%), and only one positive case (1.58%) was recorded in the age group less than 1year. The highest infection rate of B. besnoiti infection was recorded in the female animals (14.95%) followed by the male animals (8.33). This is the first report on the detection of B. besnoiti in cattle and water buffaloes in Egypt.

Blood-feeding patterns of horse flies in the French Pyrenees

Horse flies can mechanically transmit Besnoitia besnoiti, the agent of bovine besnoitiosis. Although previously limited to enzootic areas, especially the French Pyrenees Mountains, bovine besnoitiosis is now considered a re-emerging disease in western Europe. To improve understanding of the role of horse flies as mechanical vectors, this study investigated their blood-feeding ecology in the eastern French Pyrenees, in two high-altitude summer pastures whose main domestic ungulates were cattle, and in a wildlife park with native fauna. Species-specific PCR assays were conducted to identify the sources of blood meals: wild boar, horse, cattle (or bison), sheep (or mouflon), goat, red deer, roe deer and izard (or Pyrenean chamois). In La Mouline pasture, tabanids (N=20) fed on red deer (70%) and cattle (30%). In Mantet pasture, tabanids (N=24) fed on cattle (52%), red deer (20%), wild boar (16%), horse (8%) and sheep (4%). In the wildlife park, Tabanus bromius (N=32), the most abundant species collected, fed on red deer (85%), bison (9%) and wild boar (6%). Despite relatively high densities in both the pastures and in the wildlife park, small wild ungulates (izard, mouflon and roe deer) were not detected as a source of blood meals. Only two mixed blood meals were identified in two specimens of T. bromius: cattle/horse for the specimen collected in the pastures, and bison/wild boar for the specimen collected in the wildlife park. Our findings showed that tabanids display a level of opportunistic feeding behaviour, in addition to a preference for red deer, the latter being particularly true for Philipomyia aprica, the most abundant species collected in the pastures.

Bovine besnoitiosis in Switzerland: Imported cases and local transmission

Bovine besnoitiosis is an economically important disease of cattle, caused by Besnoitia besnoiti (Protozoa, Apicomplexa). A considerable spreading of this parasitic infection has been observed in Europe in the last ten years, mainly related to animal trade. In order to investigate the possibility of B. besnoiti being unnoticed introduced and getting established in Switzerland through the import of breeding cattle from France, a total of 767 animals (650 cattle imported from France and 117 cattle that had contact with B. besnoiti positive cattle in Swiss farms) were screened for antibodies against B. besnoiti by both a commercial ELISA and by the indirect fluorescent antibody test (IFAT). A total of 101 (13.17%) samples showed a positive reaction in ELISA (cut-off: percent of positivity [PP] ≥15) and 16 (2.09%) samples had IFAT titers ≥ 1:100. Eight of those samples reacted positive in Western blot (WB), corresponding to five imported Limousin cattle (two cows and one bull from France and two cows from Germany) and to three cattle born in Switzerland (one Limousin heifer born from one of the positive German cows, and two adult Braunvieh cows, that had been in contact with one of the French cows at a Swiss farm). Seven of those animals were subclinically infected and one animal showed only very mild signs. They were subsequently slaughtered, and the serological diagnosis could be confirmed by real-time PCR and/or histopathology in seven animals. The most frequent parasite localizations were the tendons and surrounding connective tissue of the distal limbs and the skin of the head region. Furthermore, B. besnoiti could be successfully isolated in vitro from one French, one German and one Swiss cattle (isolates Bb-IPZ-1-CH, Bb-IPZ-2-CH and Bb-IPZ-3-CH). In the current situation in Switzerland, prophylactic and control measures should include a serological examination of cattle to be imported from endemic areas and the culling of all confirmed positive animals from the herd. The evidence of B. besnoiti infection in both imported and locally born cattle shows that the conditions for the establishment and dissemination of this parasite in Switzerland seem to be adequate.