Three cases of ophthalmomyiasis externa by sheep botfly Oestrus ovis in Italy

Human infection with the sheep nasal botfly Oestrus ovis is sporadic and is often the consequence of an accidental deposit of the larvae by an adult botfly in the eye. This infestation results in external ophthalmomyiasis that, although a very rare condition, is more common among people living close to farming communities. We report three cases of O. ovis infestation which occurred in Italy in a limited area of La Spezia province (Le Cinque Terre), Italy during summer 2004. None of the patients had contact with wild or farm animals.     

Serum and skin surface antibodies and their associations with sheep biting lice, Bovicola ovis, on experimentally infested sheep

The sheep biting louse ( Bovicola ovis ) feeds superficially on the skin of sheep but appears to stimulate an immune response. In this study we examined the association between louse infestation and serum and skin surface antibodies. Louse numbers were monitored on experimentally infested Polypay and Columbia ewes for two years and on their lambs in the second year. Serum and skin wash samples were tested for antibodies to soluble extracts of B. ovis , Stomoxys calcitrans and Musca autumnali s by enzyme linked immunosorbent assay (ELISA). In addition, the effects of skin wash extracts on B. ovis were examined in vitro .
The titre of anti- B. ovis antibodies in the serum did not differ significantly between infested and naive ewes. However, there was an increase in serum antibody titre which coincided with periods of high louse density in ewes with high louse counts. Infested lambs had higher serum antibody levels than naive lambs. Substantial cross reactivity was evident among extracts of the different insects.
Densities of lice on the ewes during population decline were negatively related to the titre of skin surface antibodies. Skin washings collected from sheep during B. ovis population decline reduced the number of louse progeny when incorporated into louse diet. These results indicate that B. ovis stimulates an immune response in sheep and suggest that compounds on the skin surface may play a role in the regulation of louse populations.     

A physiological and biochemical model for digestion in the ectoparasitic mite,Psoroptes ovis (Acari: Psoroptidae)

Mites are an important group of arthropod pests affecting crops, animals and humans. Despite this, detailed physiological studies on these organisms remain sparse due largely to their small size. Unifying models are required to draw together the diverse information from studies on different groups and species. This paper describes a model for digestion in the parasitic mite, Psoroptes ovis, the causative agent of psoroptic mange or sheep scab disease. The limited information about this species is supplemented with data from other acarines, especially house dust mites and ticks. We review the range of enzymes and allergens found in mites and consider their possible roles in digestion in mites, generally and in particular, P. ovis. Histological studies, enzyme biochemistry and molecular biology and experimental evidence suggest that P. ovis utilises a digestive system reliant upon acid peptidases functioning in a largely intracellular environment. The actions of the digestive enzymes are supplemented by the involvement of bacteria as potential direct and indirect sources of nutrition. It is possible that some extra-corporeal digestion also takes place. The interaction of bacteria and digestive enzymes on the skin surface of the sheep may be responsible for the excessive pathological reactions evident in clinical sheep scab.     

Hypoderma lineatum antigen and anti-Przhevalskiana silenus antibodies: cross-reactivity and antibody kinetics in naturally infested goats.

To evaluate the cross-reactivity between Hypoderma lineatum antigen and anti-Przhevalskiana silenus antibodies six protocols with different concentrations of antigen and different dilutions of sera and conjugate were applied. The highest cross-reaction between the H. lineatum antigen and the anti-P. silenus antibodies is given by 2 micrograms/ml of antigen concentration, 1:400 of serum and 1:10,000 conjugate dilution. The study on the kinetic development of antibodies in goats naturally infested by P. silenus and the natural course of infestation pointed out the existence of a good correlation between individual antibody kinetics and the natural evolution of the cycle of infestation. The highest antibody concentration may be registered from October through November, coinciding with the end of the migration fo the larvae inside the animal's body. In our condition, this period can be considered as a favorable sampling period for immunodiagnosis and immunoepidemiological studies of goat warble fly infestation.     

Effect of infestation with Psoroptes ovis on the nocturnal rubbing and lying behaviour of housed sheep

The relationship between Psoroptes ovis infestation and the nocturnal rubbing and lying behaviour of sheep was examined as part of a longitudinal study of sheep scab. A total of 40 non-infested, scab-na?ve sheep were divided into six groups of between 6 and 20 individuals and each group was subsequently penned with a single infested index case. Multilevel statistical models showed that 75% of the variation in rubbing behaviour could be explained by a combination of three variables: the age of the lesion, the size of the affected area and the time since the introduction of the index case. There were significant differences between sheep in both the baseline level of rubbing activity and the rate at which this behaviour increased over time. Increased rubbing behaviour was associated with reduced total lying times and more interrupted lying behaviour. This work has contributed to our understanding of the role of parasite-induced behavioural changes on welfare and disease transmission.     

History, biology and control of sheep scab

Sheep scab is one of the oldest known diseases of sheep. It is caused by either of the mange mites Psoroptes ovis (Fig. 1 and cover) or Sarcoptes scabiei. P. ovis causes irritation so intense that sheep become preoccupied with scratching, cease to feed and rapidly become emaciated. It should be eradicated on welfare grounds alone (Fig. 2).     

Monoclonal antibodies against acidic phosphoproteins P0, P1, and P2 of eukaryotic ribosomes as functional probes.

Mice were immunized against ribosomal acidic proteins P1 and P2 from Artemia salina, and three kinds of monoclonal antibodies were isolated. One recognized P0 in addition to both P1 and P2 (anti-P). The other two recognized either P1 (anti-P1) or P2 (anti-P2) specifically and did not recognize P0. The anti-P antibody, but not anti-P1 or anti-P2, recognized a 22-amino acid peptide corresponding to the carboxyl-terminal sequence common to P1 and P2. This antibody, but not the others, inhibited poly(U)-directed polyphenylalanine synthesis. The anti-P1 bound to ribosomes but failed to inhibit polyphenylalanine synthesis: the anti-P2 did not bind to ribosomes at all. The anti-P and its Fab fragments inhibited the elongation step of protein synthesis, namely, the binding of elongation factors 1 alpha and 2 to ribosomes as well as their ribosome-coupled GTPase activities. Anti-P had little effect on the nonenzymatic phenylalanyl-tRNA binding to ribosomes and on peptidyltransferase activity. These results suggest the functional importance of the homologous carboxyl-terminal region of the three P proteins for the interaction of the ribosome with the two elongation factors. The epitope of anti-P1 must reside in a region of the protein which is not directly involved in its function.     

Association of Maedi Visna virus with Brucella ovis infection in rams

Maedi Visna Virus (MVV) is the etiological agent of a systemic disease of sheep, which causes lesions in lungs, the central nervous system, joints, and mammary glands. It has been speculated that the association with Brucella ovis may lead to the venereal shedding of the virus. In this work, samples of epididymis from ten rams positive for MVV and infected experimentally with Brucella ovis, were subjected to liquid-phase PCR, immunohistochemistry (IHC) and in situ PCR tests, aimed at identifying the pathogens in a tissue context. IHC was carried out using a monoclonal antibody raised against p28 MVV protein and a polyclonal antibody to B. ovis. Liquid phase- and in situ PCR were designed to amplify a portion of MVV proviral DNA Pol sequence. In the animals showing B. ovis-related histopathological changes, IHC clearly demonstrated a positivity for B. ovis and MVV in interstitial and epithelial ductal cells. In situ PCR assessed the presence of MVV proviral DNA in macrophages and elements inside the epithelium. The unaffected and reagent control samples constantly gave negative results. Taken together, these data demonstrate that MVV may affect ovine epididymis, apparently taking advantage of the concurrent infection by B. ovis. The tropism of MVV for the epididymal epithelial cells, may be responsible for its excretion with the semen.     

Immunologic comparisons of Pneumocystis carinii strains obtained from rats, ferrets, and mice using convalescent sera from the same sources.

Pneumocystis carinii (Pc) infections were developed in animals immunosuppressed by dexamethasone treatment either from activation of latent infection (ferret) or trans-tracheal inoculation of Pc obtained from infected lungs of the homologous species (rat, mouse). Convalescent antisera were obtained by stopping dexamethasone treatment after 2-4 wk and allowing 5-8 wk for recovery. Parasites from infected lungs were purified by differential filtration, solubilized in loading buffer, subjected to sodium dodecyl sulfate- polyacrylamide gel electrophoresis, and blotted to polyvinylidene fluoride sheets for Western analysis. Antisera from each animal species were reacted on Western blots of antigens from rat, ferret, and mouse. Each combination of antigen and antibody from the same species of animal showed reaction with 5 or more bands of Pc antigen. Convalescent mouse antibody did not react with rat or ferret antigens. Convalescent rat antibody reacted with a mouse antigen at about 66 kDa but not with ferret antigen, and convalescent ferret antibody showed minimal, probably non-specific reactions with both rat and mouse antigens. Variations in reactions indicate antigenic differences in Pc strains infecting these animals.     

Dot-ELISA assessment of guinea pig antibody responses to repeated Dermacentor andersoni infestations

Acquired resistance to ixodid tick infestation is expressed by cattle and laboratory animals. Humoral factors appear to be involved in host acquired resistance to tick bite; however, specific immune responses have yet to be fully characterized. This study examined tick resistance expressed by Hartley guinea pigs upon repeated infestation with Dermacentor andersoni, and describes longitudinal development of antigen specific immunoglobulin over approximately 180 days. Guinea pigs were infested either 4 times with D. andersoni adults, or twice with nymphs. Both infestation groups, adults and nymphs, demonstrated a significant level of resistance to re-infestation, following initial exposure. Dot enzyme-linked immunosorbent assay (Dot-ELISA) was employed to detect antibody reactive with D. andersoni salivary gland antigens (SGA). Animals infested with adults had antibody that increased at a relatively constant rate until the fourth infestation, which was differentiated by a sharp increase in titer, that was maintained for approximately 2 wk. Guinea pigs that received nymph infestations had SGA-specific antibody; however, titers were lower than those in the adult infestation group. Antibody levels continued to increase approximately 80 days beyond the final (second) infestation for this group. A direct correlation between resistance and antibody titer was not evident, since resistance was relatively stable after the second infestation in both infestation groups, and tick-specific immunoglobulin levels continually increased.     

Immune failure in the absence of profound CD4+ T-lymphocyte depletion in simian immunodeficiency virus-infected rapid progressor macaques

A fraction of simian immunodeficiency virus (SIV)-infected macaques develop rapidly progressive disease in the apparent absence of detectable SIV-specific antibody responses. To characterize the immunopathogenesis of this syndrome, we studied viral load, CD4+ T-lymphocyte numbers as well as cellular and humoral immune responses to SIV and other exogenous antigens in four SIVsm-infected rhesus macaques that progressed to AIDS 9 to 16 weeks postinoculation. Each of these animals exhibited high levels of viremia but showed relatively preserved CD4 T lymphocytes in blood and lymphoid tissues at the time of death. Transient SIV-specific antibody responses and cytotoxic T-lymphocyte responses were observed at 2 to 4 weeks postinoculation. Two of the macaques that were immunized sequentially with tetanus toxoid and hepatitis A virus failed to develop antibody to either antigen. These studies show that the SIV-infected rapid progressor macaques initially mounted an appropriate but transient cellular and humoral immune response. The subsequent immune defect in these animals appeared to be global, affecting both cellular and humoral immunity to SIV as well as immune responses against unrelated antigens. The lack of CD4 depletion and loss of humoral and cellular immune responses suggest that their immune defect may be due to an early loss in T helper function.     

Prevalence and intensity of Oestrus ovis in Akkaraman sheep in the Konya region of Turkey

Slaughterhouse surveys to determine the prevalence and intensity of larval Oestrus ovis Linnaeus (Diptera: Oestridae) in sheep, were conducted monthly for 1 year in Konya, Turkey. A total of 624 sheep, selected at random, were examined and 59% were found to be infested by O. ovis. A total of 8801 larvae were collected, of which 68.9% were first-stage, 19.1% second-stage and 12% third-stage larvae. All three larval stadia were seen in each month of the year. The larval intensity for infected sheep was 23.9, with 16.48 L(1), 4.55 L(2) and 2.87 L(3). The monthly prevalence ranged from 34.6% in January to 76.9% in October. The largest number of larvae (180) was obtained from a sheep in August (122 L(1), 52 L(2) and 6 L(3)). The infestation rate was higher in 4 - 6-year-old sheep, at 72.6%. The infestation rates were 64.4% in female and 47.5% in male sheep.     

Analysis of larval antigens of Oestrus ovis for the diagnosis of oestrosis by enzyme-linked immunosorbent assay

A serodiagnostic test for the diagnosis of infestation by the sheep nasal bot fly, Oestrus ovis (Linné) was examined. The enzyme-linked immunosorbent assay (ELISA) technique was used to analyze and compare the production of immunoglobulin G (IgG) antibodies against excretory-secretory products (ESP) and crude extract (CE) antigens from all the different larval stages of O. ovis in the sera of 276 adult sheep sampled in summer (n = 135) and winter (n = 141). ESP from first stage larvae was the most sensitive, coating antigen in winter and ESP from second stage larvae during summer. The most specific values were obtained by ESP against L1 in winter and by CE against L3 in summer. These results show that the stage of larval development has a significant impact on the humoral immune response over the course of a season. A significant correlation (P < 0.001) was found between the number of O. ovis larvae and the serum antibody levels using all differents antigens, except L3 CE. In Spain, where a long favourable period exists for the evolution and development of the different stage larvae between March and November, the ELISA test using L1 ESP antigen during winter and L2 ESP antigen in summer may be used for ovine oestrosis immunodiagnosis.     

In vitro oncosphere-killing assays to determine immunity to the larvae of Taenia pisiformis, Taenia ovis, Taenia saginata, and Taenia solium

Taeniid cestodes infect humans and livestock, causing considerable morbidity and mortality, as well as economic loss. Substantial progress has been made toward the production of recombinant vaccines against cysticercosis in livestock animals. Further development of these vaccines would be aided if a reliable in vitro test were available to measure host-protective immune responses in vaccinated animals. Here, we describe in vitro oncosphere-killing assays for the quantification of host-protective serum antibodies against Taenia pisiformis, Taenia ovis, Taenia saginata, and Taenia solium in rabbits, sheep, cattle, and pigs, respectively. Activated oncospheres of T. pisiformis, T. ovis, T. saginata, and T. solium were incubated in vitro in culture medium, test serum, and a source of complement, and oncosphere killing was assessed after 10 days of culture. In vitro oncosphere killing reflected the presence of specific antibody, and the oncosphere-killing assay typically indicated immunity to the homologous parasite that had been determined in vivo. This study describes the first reliable oncosphere-killing assays for T. pisiformis, T. ovis, T. saginata, and T. solium. These assays will be used for further research into the optimization of recombinant vaccines against cysticercosis.     

Sheep and goat immune responses to nose bot infestation: a review

Oestrus ovis L. (Diptera: Oestridae) is a cosmopolitan agent of myiasis in sheep and goats. The parasitic phase begins after adult females deposit first-stage larvae (L1) into the nostrils of hosts; these larvae develop into L2 and L3 in the nasal and sinus horn cavities. Sneezing and nasal discharges are the major clinical signs in infected animals. The pathogenesis of O. ovis infection is caused by: (a) the trauma resulting from the mechanical action of spines and hooks during larval movement on mucosal membranes, and, more importantly, (b) an allergenic reaction provoked by molecules excreted/secreted by larvae, of which salivary antigens are those mainly recognized by the host's immune system. The recruitment of immune reactive cells increases gradually from the nasal to sinus cavities in infected hosts. Mast cells, eosinophils, macrophages and lymphocytes are always more numerous in infected than non-infected animals. Humoral (antibody) systemic response of immunoglobulin G (IgG) usually reaches seroconversion 2-4 weeks post-first infection and the highest levels are observed during the development of L2 and L3 larvae. Local antibody responses include specific IgG, which has been found to negatively correlate with larval survival and development. Hypersensitivity reaction, immunomodulation, immunization trials and mixed infections of O. ovis and helminths are discussed.     

Experimental Brucella ovis infection in mouflon (Ovis musimon)

Brucella ovis was isolated for the first time in Italy in 1994 from the genital organs of two domestic rams. In subsequent years bacteriologic and serologic investigations demonstrated an increasing distribution of this disease in domestic sheep. Mouflon (Ovis musimon) occur in several hilly and mountainous areas of Italy where they can potentially contact domestic sheep. To determine if this species may have a role in the epidemiology of B. ovis, four male and four female mouflon, serologically negative for B. ovis and other Brucella spp., were infected intra-conjunctivally with B. ovis strain BG1/94. Physical examinations, including collection of blood samples for serology and bacteriology, were performed weekly. The animals were euthanized 8 mo postinoculation (p.i.). Samples of retropharyngeal, parotid, and iliac lymph nodes; bone marrow; kidneys; spleen; epididymis; testicle; bulbourethral glands; seminal vesicles; uterus; and oviducts were collected from each animal as appropriate for histopathology and bacteriology. At the time of euthanasia none of the animals exhibited obvious clinical signs of brucellosis. The animals seroconverted 2 wk p.i. and became seronegative 24 wk p.i. Bacterial cultures, including hemocultures, were negative. No lesions due to B. ovis infection were revealed by histologic examinations. Brucella ovis probably did not infect mouflon and this wild sheep is not likely to play a role in the epidemiology of contagious epididymitis caused by B. ovis.     

The influence of supplemental vitamins A and E on ovine humoral immune response

These experiments determined if supplemental vitamins A and/or E would enhance ovine antibody responses. All-rac-alpha-tocopheryl acetate was fed to lambs approximately 6 months old (30 to 40 kg) at levels of 33 (controls), 121, 276, 396, and 476 mg/kg of feed (which are total vitamin E levels). Primary and secondary immunizations with 10 mg keyhole limpet hemocyanin (KLH) were given. A nonlinear dose response of serum antibody titers was observed and the 476 mg vitamin E/kg treatment significantly enhanced (P less than 0.05) the peak primary response over controls. Retinyl acetate fed at five levels ranging from 7000 (the control level) to 97,400 IU/kg feed failed to influence antibody production to 10 mg KLH of lambs about 6 months old (29 to 41 kg). There was no detectable response to an ovalbumin antigen (100 mg). Neonatal lambs were injected with retinyl palmitate or the carrier of the injected vitamin. These lambs failed to raise antibody titers to either of the antigens administered (10 mg KLH, 100 mg ovalbumin). This was apparently due to a neonatal period of immune paralysis to certain antigens. A preliminary study showed that no KLH-specific antibodies are detectable in lambs immunized earlier than 7 weeks of age. Lambs in this age range were utilized in the last trial in which four treatments were applied: 3000 mg oral vitamin E, 400,000 IU injected vitamin A, 4 ml of the injectable vitamin A carrier, or no treatment. Half of the animals in each of these groups were immunized with 15 mg KLH and 1 ml Brucella ovis bacterin and the other half served as nonimmunized controls. No significant differences in titers to KLH were observed. Lambs receiving 3000 mg vitamin E or the carrier produced secondary peak anti-B. ovis titers higher (P less than 0.05) than those of the untreated controls.     

The duration of passive protection against Taenia ovis larvae in lambs

In an attempt to induce passive protection in lambs against Taenia ovis larvae that would last for the 15-20 weeks from birth to slaughter as fat lambs, one group of ewes was immunized by a series of injections of 2000, 4000, 8000, 16 000 and 32 000 activated oncospheres of Taenia ovis prior to parturition. Another group of ewes was not immunized. All ewes had previously grazed pasture lightly infected with T. ovis eggs. Most lambs from non-immunized ewes developed cysts after oral infection with T. ovis eggs. However, no lambs from immunized ewes developed cysts up to and including 6 weeks after birth. Between 8 and 16 weeks after birth a proportion of lambs were found to be susceptible to infection. By 18 weeks after birth all lambs were apparently susceptible. The 99% confidence band for the mean duration of demonstrable complement-fixing antibody titres was 6.2-7.8 weeks for lambs from immunized ewes. The persistence of maternal protective antibody in some lambs could possibly preclude successful active immunization of all lambs against T. ovis larvae before 18 weeks of age.     

Transfer of experimental allergic neuritis with P2-reactive T-cell lines

Experimental allergic neuritis (EAN) was induced in normal Lewis rats by systemic passive transfer of T-cell lines responding to P2 protein. These cells had predominantly helper phenotype and could induce EAN within 7 days following adoptive transfer. There was no anti-P2 antibody response in the recipients of the P2-reactive cells recovered from donors with high anti-P2 antibody levels. This study provides direct evidence that T cells are important for the induction of EAN. Furthermore, there was no evidence of a pathogenic role for anti-P2 antibody in passive EAN.