The identification of a VDAC-like protein involved in the interaction of Babesia bigemina sexual stages with Rhipicephalus microplus midgut cells

Oestrosis is highly prevalent in Mediterranean countries. Understanding the life cycle of Oestrus ovis is crucial to design effective control measures of this myiasis, largely based on the use of macrocyclic lactones. We carried out a survey of ovine oestrosis in the island of Majorca (Spain) and found that 46.03% of animals were infested in a 13-month period. Interestingly, we found significant differences in oestrosis prevalences in winter and autumn when separating the animals by group of age (P<0.001). Pearson correlation analysis showed that prevalence in lambs younger than 4 months was significantly affected by changes in air temperature (P<0.05), but this association was not significant in adult sheep (P=0.081). Chronic infestations or unsystematic treatments may explain confusing results in adult sheep. Observing the evolution of the disease in young lambs, we determined that the hypobiotic period took place from October to February and the beginning of fly activity occurred between May and June. Interannual variations in oestrosis prevalence indicate the need of monitoring the disease to establish the appropriate timing of treatments. We hypothesize that lambs are better indicators of the seasonality of oestrosis than their older counterparts. Furthermore, we propose that observing O. ovis infestations in young lambs can be used as an efficient early warning system of fly activity, to be applied in future control programs.     

Comparison of infection rates of Oestrus ovis between sheep and goats kept in mixed flocks

Oestrosis is a nasal myiasis of sheep and goats caused by larvae of the fly Oestrus ovis and can lead to severe clinical signs, which together with the disturbance caused by the adult fly may result into serious economic losses. Infection rates and larval burdens are always higher in sheep than in goats after either natural or artificial infestation. The aim of this study was to compare the host preference of the adult fly O. ovis between sheep and goats in mixed flocks, where they are kept together under the same husbandry conditions and hence, are very similarly exposed to the fly preference. Blood sera samples were collected from a total of 397 sheep and 335 goats, from 43 mixed flocks located at different regions of Greece. Antibodies specific to O. ovis IgG were measured by ELISA. A flock was considered positive when at least one individual was positive, i.e. showed a seropositivity of >or=20% in relation to positive control sera. A total of 193 (48.6%) sheep and 58 (17.9%) goats were found to be seropositive against O. ovis. Thirty-eight (88.4%) out of 43 flocks had at least one seropositive animal. The mean seroconversion against O. ovis in animals from the different flocks was 38.6% and 13.6% for sheep and goats, respectively, whereas the variance of infection within each flock was 0-100%. The mean seropositivity between sheep that were found to be positive or negative was 60.6% and 5.4%, respectively, whereas the corresponding values between goats were 35.2% and 5.2%, respectively. No significant difference in the seroconversion values was noted between flocks from the different areas (P=0.817), whereas a very significant difference was observed between animal species (P=0.001). However, there was no significant difference when seroconversion comparisons were made within samples of the same animals species, sheep or goats from different flocks of all the regions included in the study (P=0.695). The results of this study clearly demonstrate that O. ovis has a widespread distribution in Greece, and the seroprevalence is significantly higher in sheep than goats (P=0.001).     

Analysis of the humoral immune response to Oestrus ovis in ovine

Antibody responses (IgG, IgM and IgA) against Oestrus ovis were analyzed in sheep and in first year grazing lambs from Sardinia (Italy) by an indirect-enzyme-linked immunoassay test and L2 O. ovis excretory/secretory antigens. Serum samples from 208 sheep were obtained prior to be slaughtered, and then heads were removed and cut open along their longitudinal axis to collect the parasites from the nasal cavities, turbinates and sinus. Besides this, blood samples were monthly collected from the lambs of G-1 (maintained under field conditions) and the lambs of G-2 (kept housed since birth to avoid Oestrus infestations) throughout a year. In the sheep, a positive significant correlation was observed between the number of first instar O. ovis larvae and the values of IgM, and between the second instar larvae and the IgG optical densities. In the lambs, all classes of antibodies increased significantly from July in G-1. The highest values of IgG were reached in September (IgG) and decreased in November-December. The IgM response peaked in November, and very low values of IgA were observed during the study. Matching these data with chronobiology of O. ovis in this region, we conclude that the first infection occurs on May, stimulating the production of humoral antibodies. The reduction of the IgG antibody levels starting from October means the beginning of the diapause while the IgM response seems to be associated to the presence of L1 in the nasal cavities. The data obtained led us to forecast an early treatment of the ovine on June-July, which should keep away from the maturation of O. ovis L1 larvae, avoiding the development of clinical lesions and interrupting the life cycle of this parasite.     

Seasonal variation of Oestrus ovis-specific antibodies in sheep and goats mixed flocks in Greece

The aim of this survey was to investigate the year-round epidemiological patterns of Oestrus ovis ELISA sero-prevalence in sheep and goats kept together under the same husbandry system in an endemic area of Greece. Twenty-five adult female sheep and 25 adult female goats, coming from a large mixed flock, were randomly selected, eartaged and monthly blood sampled during 1 year period (November 1998-October 1999). Serological prevalence in sheep was 100% all around the year. Mean intensities of specific O. ovis antibodies follow a seasonal evolution with higher mean titers between March and July than in winter. In contrast, the serological prevalences in goats were low specially in winter months (from October to January). No significant difference were noticed in goats antibody levels during the year period. The possible reasons of this difference of O. ovis sero-prevalence between sheep and goats are discussed.     

Serine protease activity in excretory-secretory products of Oestrus ovis (Diptera: Oestridae) larvae

The sheep bot fly, Oestrus ovis, is a very common myiasis of nasal and sinus cavities of sheep and goats causing severe welfare and production implications. As the viability of O. ovis adult flies strictly depends on larval abilities to assimilate and to stock nutrients from the host, it was necessary to investigate proteolytic activities in larval excretory/secretory products (ESP). ESP of O. ovis larvae degrade mucosal and plasmatic components such as mucin, albumin or immunoglobulin G. A preliminary biochemical characterization, using substrate gel analysis and inhibitor sensitivity, demonstrated the presence of at least six major serine proteases (molecular weights from 20 to 100 kDa), mainly trypsin-like, secreted in the digestive tube of larvae. Their involvement in larval trophic activity and evasion from the host immune response is further discussed as O. ovis excretory/secretory serine proteases could represent potential vaccinal targets.     

Evaluation of the conjunctival swab for canine visceral leishmaniasis diagnosis by PCR-hybridization in Minas Gerais State, Brazil

Larvae of Oestrus ovis (Diptera: Oestridae) are ubiquitous parasites of nasal and sinusal cavities of sheep and goats. According to the chronobiology of O. ovis infections in Sardinia and the seasonal pattern of the IgG response, the optimal period to investigate the relationships between O. ovis larval populations and intensity of local and systemic IgG antibody responses was mid-July in the summer season. Sarda x Lacaune ewes (n=186), divided into three ram-families were used in the study. Systemic and local IgG responses were measured by ELISA tests using second stage larval crude extracts (L2CE) and L2 (L2SGC) and L3 (L3SGC) salivary gland contents as coating antigens. The number of larval instars, larval length of L1, L2 and L3 larvae, and larval weight of L2 and L3 larvae were individually recorded after ewe necropsy. Negative correlations among larval establishment and/or larval development on the one hand and intensity of local or systemic IgG responses on the other hand were found in two out of three studied ram-families.     

Concurrent parasitic infections of sheep: depression of Trichostrongylus colubriformis populations by a subsequent infection with Oestrus ovis

Concurrent infections of sheep with Oestrus ovis and trichostrongyles of the digestive tract are common in the field. Previous results have shown that a previous infection with O. ovis adversely affects worm populations of either Trichostrongylus colubriformis or Haemonchus contortus. However, no information was available to determine the influence of the succession of infections on the expression of interactions between these parasites located in remote anatomical sites. In order to investigate the role of these modulating factors, an experimental study was conducted on four groups of na?ve sheep, examining the consequences of a delayed infection with O. ovis on a pre-existing population of T. colubriformis. group T was infected four times with 4000 T. colubriformis larvae on days 0, 14, 28 and 42 of experiment; group O received multiple infections with O. ovis first instar larvae on days 42, 49, 56, 70 and 77; sheep from group TO received both infections and animals from group C remained as uninfected controls. Faecal egg counts and eosinophilia were measured weekly throughout the study. At necropsy (day 91), the mucosal cellular responses in the nasal cavities (septum, turbinates, ethmoid and sinus) and in the digestive tract (stomach and small intestine) from all animals were analysed from histological sections. Infection of the digestive tract with nematodes did not modify the biology of Oestrus populations, as measured by the number and weight of larvae. In contrast, infections with O. ovis after T. colubriformis infection was related to significant reductions (P < 0.01) in nematode egg excretion and worm burdens. These changes were associated with significant modifications in populations of mast cells, globule leucocytes and eosinophils in the respiratory and digestive tracts. These results indicate that an antagonistic interaction exists between the populations of O. ovis in the nasal cavities and T. colubriformis in the small intestine but that the order of succession of infections with the two parasites is not a major modulating factor for expression of interactions. They also confirm that parasitic infection in one particular anatomical site induces "at distance" inflammatory reactions of the whole mucosal system.     

Regulation of Oestrus ovis (Diptera: Oestridae) populations in previously exposed and naïve sheep

Larvae of Oestrus ovis (Insecta: Diptera: Oestridae) are common parasites of nasal and sinus cavities of sheep and goats. During larval development, a specific immune reaction is initiated by the host with a humoral local and systemic response and the recruitment of eosinophils and mast cells in the upper airways mucosae. Nevertheless, the roles of these responses in the regulation of O. ovis larvae populations in sheep are not yet known. The aim of this study was to compare the establishment and the development of larvae as well as some inflammatory or immune parameters between different groups of half-sibling sheep: (i) a primed group experimentally infected twice before a challenge infection, (ii and iii) two groups infected once only and previously treated with a long-lasting cortico?d before the challenge (one group) or not (the other group). A fourth group of non-infected animals was added in the experimental design. The larval establishment rate was 23% in the cortico?d treated group compared to about 10% in the two other infected groups. Moreover, the larval development appeared more rapid in the cortico?d treated group than in the two other infected groups suggesting that the inflammatory response is involved in the regulation of O. ovis populations. By contrast, no differences in the establishment rates were shown in the primed group compared to the na?ve group (without cortico?d treatment) despite evidence of higher eosinophilia, serum specific IgG, and immediate hypersensibility to excretory-secretory products of larvae. The specific lymphocyte proliferation was reduced in the primed group compared to the na?ve one suggesting that an immuno-suppression occurs following repetitive O. ovis infections.     

Sheep oestrosis (Oestrus ovis Linné 1761, Diptera: Oestridae) in Sardinia, Italy.

Oestrosis, the nasal myiasis of sheep and goats, is caused by the larvae of Oestrus ovis L. 1758 (Diptera, Oestridae) that develop from the first to the third stage larva in the nasal cavities and frontal sinuses of affected animals. The authors report the results of an epidemiological study of oestrosis of sheep in Sardinia, Italy. Heads of 6-month to 10-year-old Sardinian sheep (n=566) from 124 free-ranging flocks were examined for the presence and location O. ovis larvae from December 1996 to November 1997. Larvae were collected, counted, and larval stages were identified. O. ovis larvae were found in 100% of examined flocks and in 91% (514/566) of examined sheep. The monthly prevalence ranged from 69% in May to 100% in July. First stage larvae were found in 82% (463) of all heads examined, second stage larvae in 65% (367) and third stage larvae in 10% (56). The majority of sheep harboured first stage larvae, with prevalences of over 80% throughout most of the study period. The prevalence of O. ovis found in this study of Sardinian sheep is the highest reported in the Mediterranean area. The high percentage of first stage larvae found throughout the entire study period may be due to a brief period of decreased rate of larval maturation, in particular in December 1996 (96%) and January-October 1997 (94%). Third stage larvae were consistently present, often however, with extremely low prevalences compared to total larval burden.     

Parasitism by Oestrus ovis: influence of sheep breed and nematode infections

Previous studies showed that Santa Ines (SI) hair sheep were more resistant to gastrointestinal nematode infections (GIN) than Ile de France (IF) sheep. The present experiment aimed to evaluate if that reported resistance difference against GIN also occurred against Oestrus ovis infestation and also to evaluate the influence of O. ovis infestation on the gastrointestinal nematodes (GIN) infections. SI (n=12) and IF (n=12) young male lambs were weaned at 2 months of age and moved to a paddock (0.3 ha) with Brachiaria decumbens grass, where they also received concentrate ration. The animals were kept together during the experimental period (September to early December 2009). Fecal and blood samples were taken from all animals every 2 weeks and body weight and nasal discharge score (oestrosis clinic signs) were recorded on the same occasion. In early December 2009, all lambs were sacrificed and O. ovis larvae and GIN were recovered, counted and identified according to the larval stage. All animals were infested by different larval instars of O. ovis without any statistical difference between breeds (P>0.05). The SI lambs had an average of 24.8 larvae, and the intensity of infection ranged between 14 and 39 larvae, while the IF lambs showed an average of 23.5 larvae with the minimum and maximum from 11 to 36 larvae, respectively. SI lambs presented the lowest nematode fecal egg counts (FECs) and the lowest mean numbers of Haemonchus contortus, Trichostrongylus colubriformis and Strongyloides papillosus, however, there was no significant differences between group means (P>0.05). Inverse relationship between numbers of O. ovis larvae and gastrointestinal nematodes was observed in both breeds. SI sheep showed a significant increase in blood eosinophils and total IgE serum levels and these variables were negatively correlated with nematode FEC. A negative correlation was observed between total IgE serum level and H. contortus burden in both breeds. In conclusion, there was no breed difference regarding O. ovis infestation and in each breed, animals with more nasal bot fly larvae tended to display smaller worm burden.     

Prevalence and larval burden of oestrus ovis (Linné 1761) in sheep and goats in northern mediterranean region of France

A slaughterhouse survey to determine prevalence and larval burden of Oestrus ovis larvae in sheep and goats was performed monthly during one year in Pézenas, South of France, northern mediterranean region. A total of 1303 sheep and goat heads were selected at random. O. ovis larvae were found in 274 sheep out of 631 (43.4%), and the prevalence rate varied from 14.3% in February to 65% in October. The mean number of larvae in infected sheep heads was 10.86 with 9.24 L1, 0.91 L2 and 0.71 L3. One hundred and ninety-one goats out of 672 were infected (28.4%), and the prevalence rate varied from 6.25% in September to 47.1% in April. In infected goat heads, the mean parasitic burden was 5.35 with 4.04 L1, 0.73 L2 and 0.58 L3. These results confirm worldwide observations indicating that the prevalence and the parasitic burdens are less in goats than in sheep.     

Genetic structure of Oestrus ovis populations in sheep and goats.

A genetic analysis using RAPD markers was performed on 12 natural populations of Oestrus ovis (Linné, 1761). Three-hundred and six O. ovis larvae (first, second and third instars) were randomly recovered in nasal cavities of sheep and goats naturally infected in Algeria, Ethiopia, France, Mauritania, Rumania and Tunisia and were analysed by 56 RAPD fragments. The results showed a high diversity within all samples. A significant genetic divergence was showed by discriminant analyses among the 12 populations sampled (p<0.0001). Moreover, discriminant analyses showed significant differentiation (p<0.0001) between O. ovis larva populations of sheep and goats and also among samples collected in the same region.     

Evaluation of a direct ELISA for the serodiagnosis of Oestrus ovis infections in sheep.

Oestrosis is a parasitic disease of sheep and goats caused by the nasal bot fly Oestrus ovis. In the United Kingdom the economic losses as a result of infestation can be considered negligible, but the differentiation of O ovis cases from more serious diseases such as listeriosis, gid and sheep scab is of considerable importance. Currently, diagnosis of oestrosis relies on the subjective observation of clinical signs or the demonstration of larvae postmortem. This paper assesses the effectiveness of a direct enzyme-linked immunosorbent assay (ELISA) using a crude somatic antigen from first-stage larvae (L1) in the serodiagnosis of oestrosis. The system has been validated with sera from both endemic and non-endemic areas and the results correlated with the clinical data found postmortem. The sensitivity and specificity of the assay were 97.4 per cent and 97.6 per cent, respectively, using a cut-off point based on 35 per cent binding of a reference positive control serum.     

Incidence of Oestrus ovis infestation in Borno-white Sahel goats in the semi-arid zone of Nigeria

Research conducted on the incidence of Oestrus ovis in Maiduguri showed that 53.8% of the 4,000 Borno-White Sahel goats examined were parasitised. The mean year-round infestation per goat was 2.03 larvae. Adult goats had a higher infestation rate than younger ones (60 versus 47.5%). Infestation rates for the females and males were 55.2 and 52.2%, respectively. Nevertheless there was no statistical significance between values for the different age and sex groups studied. Different larval stages were encountered throughout the year with a 3rd instar peak between May and September and a first instar peak from June to October suggesting a fly period between June and October.     

Oestrus ovis larval myiasis among goats in northern Jordan

From December 1998 to December 1999, heads of 520 local goats slaughtered at the Irbid, Ramtha and Howarra Abattoirs (northern Jordan) were examined for the three larval instars (L(1)-L(3)) of Oestrus ovis. Of 520 heads, 126 (24%) were infested with O. ovis larvae. All three larval instars were observed in both sexes; all age groups were infested in each month of the year. The mean age of the goats sampled was 1.5 years. The numbers of parasites infesting hosts showed a significant (P<0.05) correlation with sheep age (r(sp)=0.31-0.42) for all three larval instars. The numbers of larvae in each host followed an overdispersed distribution, which fit a negative-binomial model (but not a Poisson distribution). There were more parasites recorded in the presence of purulent discharge or laryngitis, fewer in the presence of catarrhal discharge and no association with pharyngitis sinusitis, or rhinitis.     

Oestriasis in small ruminants in Senegal. Preliminary note

Observations made in Dakar abattoirs from March to December 1987 revealed that 46.39 per cent sheep and 57.89 per cent goats carried Oestrus ovis larvae. A clear relationship between cause and effect exists between nasal discharge and the presence of mature larvae. Because of their spine covering, they are irritating enough to provoke nasal mucosa inflammation with possible microbic complications. It is thus necessary to take into account the frequency and the pathogenicity of Oestrus ovis infection in the aetiologic study of respiratory affections of the small ruminants in Senegal.     

Experimental concurrent infection of sheep with Oestrus ovis and Trichostrongylus colubriformis: negative interactions between parasite populations and related changes in the cellular responses of nasal and digestive mucosae

Concurrent infections of sheep with Oestrus ovis and trichostrongyles of the digestive tract are common in the field. In order to examine the possible occurrence of interactions between these two parasites and the consequences on parasite biology and the associated pathophysiological changes, an experimental study was conducted on four groups of na?ve sheep. Group O was infected repeatedly with O. ovis larvae from D0 to D42 of experiment; group T received a single infection of Trichostrongylus colubriformis on D49; sheep from group OT received both infections, and animals from group C remained uninfected as controls. Parasitological and pathophysiological parameters were measured regularly. At necropsy (D94), the mucosal cellular responses in the nasal cavities and digestive tract from all animals were analysed from histological sections. Infection of the digestive tract with nematodes did not modify the biology of Oestrus populations. In contrast, the presence of O. ovis was related to significant reductions in nematode egg excretion, worm fecundity and worm burdens. These changes were associated with significant modifications in tissular populations of mast cells, globule leucocytes and eosinophils in the respiratory and digestive tracts. These results indicate that parasitic infection in one particular anatomical site induces "at distance" inflammatory reactions of the whole mucosal system. The relationships between the cellular response and the changes in worm biology are discussed.     

Prevalence and seasonal incidence of larval and adult cestode infections of sheep and goats in eastern Ethiopia

A study on the prevalence and seasonal incidence of cestode parasite infections of sheep and goats was carried out in eastern Ethiopia for 2 years (May 2003-April 2005). During this period, viscera including liver, lungs, heart, kidneys and the gastro-intestinal tract were collected from 655 sheep and 632 goats slaughtered at four abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga. At the abattoirs the abdominal, thoracic and pelvic cavities as well as the muscle surfaces of all animals were visually examined for the presence of larval (cystic) stages of cestode parasites. The viscera were transported within 24 h to the parasitology laboratory of Haramaya University and were examined for larval and adult cestodes following standard procedures. The most prevalent metacestodes (larval cestodes) were Cysticercus ovis (Taenia ovis), Cysticercus tenuicollis (T. hydatigena) and hydatid cysts (Echinococcus granulosus). In sheep, the overall prevalence was 26% for C. ovis, 79% for C. tenuicollis, and 68% for hydatid cysts. Similarly, for goats, the corresponding prevalence was 22%, 53% and 65%, respectively. The difference between sheep and goats in prevalence of C. tenuicollis was significant. The high prevalence of hydatid cysts in both sheep and goats indicates that cystic echinococcosis/hydatidosis is a public health problem in these regions which requires implementation of control measures, including public health education, strict meat inspection and control of stray dogs. The results of the survey also implies that infections of small ruminants with these metacestodes are responsible for condemnation of substantial quantities of affected organs and muscles and therefore of direct economic importance. Intestinal infections with adult tapeworms of Moniezia expansa, Avitellina centripunctata and Stilesia globipunctata, and bile duct infections with Stilesia hepatica were also common in both sheep and goats. In sheep, the overall prevalence of these tapeworms were 61%, 20%, 24% and 39%, respectively. Similarly, the overall prevalence of these parasites in goats was 53%, 21%, 27% and 36%, respectively.     

Parasites of domestic and wild animals in South Africa. XXXIX. Helminth and arthropod parasites of Angora goats in the southern Karoo

Parasites were collected from 160 Angora goats and kids on the Jansenville Experimental Farm, Eastern Cape Province. Six nematodes were identified to species level and three to generic level. Nematodirus spathiger was the most numerous of the economically important nematodes recovered. It was always present, and fourth stage larvae were collected from untreated goats from August to March, while animals used as tracers also picked up most infection from August to March. Kids born on the farm during October acquired their first nematode infections between 2 and 3 months of age and the intensity of infection increased erratically thereafter to reach a plateau once the kids were 14 months of age. Eight of the nine kids between 3 and 5 months of age and examined between January and March were infected with Moniezia expansa. Five ixodid tick species were collected from the goats of which Rhipicephalus glabroscutatum was the most numerous and prevalent. Its immature stages were present mainly from March to September and adults from July to January. The goats also harboured the biting louse Damalinia limbata and the sucking louse Linognathus africanus. The greatest intensity of infestation with L. africanus occurred on the kids during the first few months of their lives. The larvae of the nasal bot fly, Oestrus ovis were present in kids at 1 month of age, and infestation took place mainly from spring to late summer.     

Efficacy of ivermectin controlled-release capsules for the control and prevention of nasal bot infestations in sheep

Objective To investigate the therapeutic and prophylactic efficacy of an ivermectin controlled-release capsule against nasal bots (Oestrus ovis) in sheep.
Design Trial 1 – A pen study with controls. Trial 2 – A field study with controls.
Animals Trial 1 – Forty Merino wethers with natural infestations of nasal bot were used. Trial 2 – One hundred nasal bot-free wethers were used.
Procedure Trial 1 – Ten randomly selected animals were slaughtered and the heads split and examined to confirm bot infestation. Fifteen animals were allocated to untreated controls and 15 to treatment with a controlled-release capsule delivering ivermectin at ≥ 20 μg/kg/day for 100 days. Twenty-nine days after treatment the sheep were killed and examined for nasal bots. Trial 2 – Nasal bot-free sheep were allocated to two groups of 45 animals. One group was untreated the other sheep were treated with capsules as above. The sheep were grazed as a single group exposed to natural challenge from O ovis . Ninety days after treatment the animals were slaughtered and examined for nasal bot infestation.
Results Trial 1 – Live O ovis larvae were recovered from 60% of control sheep. No live larvae were collected from treated sheep. Trial 2 – Forty-one percent of untreated sheep harbored nasal bot infestations. No live larvae were collected from any treated animal.
Conclusion Treatment with a single ivermectin controlled-release capsule was 100% effective against existing infestations of O ovis and as a prophylactic treatment for this parasite.