Morphological and biochemical changes in the blood of horses naturally infected with Gasterophilus sp. larvae.

Gasterophilus sp. constitute a group of specific parasites, which larval forms are found in horses and other phylogenetically related species--donkeys, mules and zebras. Their invasion is associated with marked fluctuations of hematological and biochemical blood parameters, including the activity of many enzymes and the electrolyte concentration. The purpose of the study was to analyze changes in selected biochemical and morphological blood parameters of horses infected with botfly larvae. The experimental group was formed of horses infested by Gasterophilus sp. larvae whereas the control group consisted of noninfested horses. The study was performed in two periods: at the beginning and at the most advanced invasion. Significant differences between horses at early and late stage of invasion were found in terms of erythrocyte parameters. Mean red and white blood cell counts and mean hemoglobin concentration were lower in horses with late invasion compared to those with the invasion at its early stage. The values of all the aforementioned erythrocyte parameters remained within the reference limits for both groups of horses. Moreover, both in horses infected with botfly larvae and in those from the control group, biochemical parameters studied did not exceed the respective reference limits. Our results suggest that larvae of botfly which colonize the gastrointestinal tract have relatively insignificant influence on the basic hematological and biochemical parameters of horse blood.

Experimental concurrent infection of Afar breed goats with Oestrus ovis (L1) and Haemonchus contortus (L3): interaction between parasite populations, changes in parasitological and basic haematological parameters.

The study was conducted to examine the occurrence and interaction between Oestrus ovis and Haemonchus contortus in experimentally infected Ethiopian Afar breed of goats. Twenty goats were divided into four groups (O, OH, H, and C) of five animals each. Each animal of groups O and OH received weekly infections for 5 weeks with 66 first instar larvae (L1) of O. ovis. Then animals of groups OH and H were infected with a single dose of 5000 third stage larvae (L3) of H. contortus. Goats of group C were kept free of any infection as non-infected control. Faecal egg count (FEC), blood cell count, total serum protein level and body weight were recorded weekly throughout the study period. At necropsy worm burden, female worm length, fecundity and larval burden of O. ovis in the nasal-sinus cavities of infected animals were assessed. The results showed that the presence of H. contortus in the abomasum of goats of group OH had no influence on the development of O. ovis. On the contrary, a significant reduction (P<0.05) in FEC, worm burden, fecundity and female worm length was revealed in group OH animals compared to the mono-infected animals (group H). This was associated with eosinophilia and reduced packed cell volume.

Impact of previous infestation on dynamics of circulating hypodermin C in cattle artificially infested with Hypoderma lineatum (Diptera: Oestridae).

Four groups of cattle were artificially infested with 50 first instar Hypoderma lineatum after either a primary natural infestation or in the absence of a primary infestation. In two groups the primary infestation had been terminated by the application of either an organophosphate insecticide or a macrocyclic lactone parasiticide. Circulating hypodermin C and specific antibodies were measured for 40 weeks after the artificial infestation. Stage specific mortality of the larvae was also monitored. Previously uninfested cattle exhibited typical antibody and antigen profiles during the infestations. Antibodies were first detected on Week 7p.i., they rose to maximum values between Weeks 24 and 25, then declined as larvae reached the back. A second peak occurred at Week 32 and antibody remained more less constant thereafter. Previously infested groups exhibited a dramatic anamnestic response by Week 3p.i. with antibody levels peaking at Week 8. A second peak was noted between Weeks 24 and 26p.i. after which antibody levels declined and then remained relatively stable. The dynamics of circulating hypodermin C in the previously infested cattle resembled those in the previously uninfested cattle. Mortality of first instars did not differ among the four groups. Similarly mortality of second and third instars, in the warble, did not differ although there was a tendency for higher mortality in the previously infested, untreated group. These results reinforce previous work demonstrating the development of a significant immune response during the primary infestation that is reflected in the rapid and substantial production of antibodies upon re-infestation. It is significant that a challenge model using subcutaneous injection of newly hatched first instars avoids host immune responses in the skin that result in substantial larval mortality. The current data also support the concept that migrating first instars induce significant reduction in host immune response. There is a peak of antibody production as antigen levels peak during first instar migration to the back. As larvae molt to the second instar and antigen production ceases there is a persistence of antibody which suggests release of the immune response from the suppression induced by the first instar secretory antigens.

Expression of circulating leucocytes before, during and after myiasis by Dermatobia hominis in experimentally infected rats.

Expression of circulating white blood cells was investigated in rats (Rattus norvegicus) experimentally infected with larvae of Dermatobia hominis, the human bot fly. Leucocytes were counted prior to infection (control group) as well as at 6, 10, 15, 20 and 28 days post-infection (dpi) and at 7, 15, 30 and 60 days post-larval emergence (dple). Total leucocyte numbers did not differ markedly among the groups. Significant differences were registered when values from control and animals harboring each larval stage of D. hominis were compared; with crescent rank: L1-, L2-, control and L3-infected groups. Leucocyte numbers were significantly higher in the control, 15, 20 or 28 dpi groups than in the 6 dpi animals. Higher counts were observed in control, L2- or L3-infected rats than L1-infected animals. Neutrophils, eosinophils and both large and small lymphocytes were also counted and analyzed. Basophils and monocytes were insufficient in number to permit statistical studies. These results stimulate the continuity of the studies about the host-parasite relationship in the dermatobiosis.

Some physiological responses associated with reduced wool growth during blowfly strike in Merino sheep.

OBJECTIVE: To examine the effect on wool growth of physiological changes associated with experimental flystrike in superfine Merino wethers. DESIGN: An animal house study comparing experimentally fly struck sheep with non-infected control groups that were pair fed or fed ad libitum. PROCEDURE: Sheep in the blow fly larval challenge group were each artificially infected with 500 first instar larvae per day for 8 days. Infections were terminated with insecticide on day 10. For all sheep, live weight change, feed intake and body temperature were monitored and blood collected for analysis of haematology, cortisol, serum haptoglobin, serum amyloid A and the pro-inflammatory cytokines, TNFa, IL-1 b, IL-6 and IL-8. Wool growth and staple strength were examined 4 months after infection. RESULTS: Experimental fly strike included moderate fever, depressed feed intake, and elevated cortisol, IL-6, serum amyloid A and haptoglobin. Staple strength was significantly decreased in struck sheep and did not differ between wool from sites adjacent to and remote from strike lesions. CONCLUSIONS: The results suggest that depression in feed intake alone is not sufficient to account for the decline in staple strength seen during fly strike. Cortisol, IL-6 and metabolic consequences of the acute phase response are likely to be major contributors to the systemic effects of blowfly strike on wool.

Excretion of ammonia by Lucilia cuprina larvae suppresses immunity in sheep.

Attempts to immunise sheep against natural infestations by Lucilia cuprina larvae have not been effective. Yet it is known that the larvae excrete the immunosuppressant ammonium bicarbonate. The effect of larval ammonium and nonionic ammonia on immunopathobiology was evaluated in 12 infested sheep. The concentration of ammonium in veins draining infested sites was measured in another group of four sheep. Mean jugular unionized ammonia concentration increased 3.5 to 5.6 times above pre-infested control levels. Mean venous ammonium concentrations draining infested sites were 13 times higher than pre-infested jugular or carotid levels. Increases in jugular ammonia concentrations correlated with increased number of larvae, area of infestation, earlier death, neutropenia, eosinopenia, lymphocytopenia, large declines in serum globulins and zinc, and large rises in toxic neutrophils. The high concentrations of toxic unionized ammonia in blood directly permanently damaged neutrophils and lymphocytes and depressed serum globulin production. The results show that the ammonium from the excreta of larvae of L. cuprina may be highly immunosuppressive.

Enzyme-linked immunosorbent assay using a crude antigen extract to measure Cuterebra-specific immunoglobulin G in sera of cats with systemic infections.

In cats, larvae of the dipteran fly, Cuterebra, sometimes cause severe disease by their migration through the tissues of the larynx, pharynx, nasal sinuses, brain, and spinal cord; such infected cats may die without the maggots ever reaching the subcutaneous tissues where they would typically mature. The current study examines the ability of an indirect enzyme-linked immunosorbent assay (ELISA) using crude Cuterebra antigen from maggots to detect parasite-specific immunoglobulin (Ig)G in cats with known (n = 42), suspected (n = 25), or no known exposure to the infection (n = 68). The probability of a given optical density (OD) predicting the infection status of a given animal was determined using logistic regression, and both 1:20 and 1:80 serum dilutions were highly predictive of the potential of a cat being infected with a larval Cuterebra. In 5 cases where 2 samples were collected 1-2 weeks apart, there was a mean OD increase in the second sample for both the 1:20 and 1:80 dilutions, but it was significant (P = 0.044) only at the 1:20 dilution. Sex of the sampled cat was not a significant contributor to the ability of the OD to predict the presence of a larva, but the age of the cat added significantly to the predictive value of the generated curves, with the only exception being with the 1:20 serum dilution with the curve being generated only using the cats known to be positive for larval presence. This ELISA should aid in ruling cuterebriasis in or out in suspect systemic and, specifically, neurologic cases and provide information on kinetics of antibody presence postexposure.

The investigation of lipid peroxidation, anti-oxidant levels and some hematological parameters in sheep naturally infested with Wohlfahrtia magnifica larvae.

Wohlfahrtia magnifica is the main agent of traumatic myiasis in many central and eastern European countries as well as in the Mediterranean basin and Asian countries. The present study was aimed to investigate the effect of myiasis infestation on lipid peroxidation, antioxidant levels and several hematological parameters in the sheep naturally infested with W. magnifica larvae. A total of 38 sheep, including 19 awassi sheep naturally infested with W. magnifica larvae and 19 clinically healthy awassi sheep, were studied. The infected animals were divided into three groups depending on the number of larvae (Group 1: <50 larvae; Group 2: 50-100 larvae; Group 3: >100 larvae). In blood samples, red blood cell counts, hemoglobin, hematocrit concentration and white blood cell, neutrophil, lymphocyte, eosinophil, basophil counts, plasma malondialdehyde, erythrocyte glutathione levels and erythrocyte glutathione peroxidase activity were studied. The results revealed a marked decrease in red blood cell counts and hemoglobin concentrations along with a significant increase in white blood cell and neutrophil counts. The elevation in plasma malondialdehyde levels, a function of lipid peroxidation, established a significant difference between the control group and groups 2, 3. Decreased activity of erythrocyte glutathione peroxidase was found significant in the control group and all parasitemia. We conclude that in natural infestations with W. magnifica, as in infestation with Lucilia cuprina, there is a notable increase in inflammatory activities resulting from the movements, secretions, and toxins of the larvae and form the toxins secreted by the bacteria - which leads to an impression of anemia - and that the tissue injury results in an increase in level of free radicals in the organism.

Antioxidant enzymes in erythrocytes from goats seropositive to the sheep nose bot fly (Oestrus ovis L., Diptera: Oestridae) infection.

Oestrus ovis (Diptera: Oestridae) causes an important cosmopolitan parasitosis of the nasal and sinusal cavities of sheep and goats called oestrosis. Our objective was to analyze the participation of erythrocytes in the antioxidant system in goats seropositive to O. ovis infection under field conditions. Fifty female goats naturally exposed to O. ovis infection from Baja California Sur, México, were blood-sampled. Erythrocytic intracellular content was obtained from blood plasma. Oestrosis serodiagnosis was determined by ELISA. Protein, hemoglobin (Hb), superoxide dismutase (SOD), mieloperoxidase (MPO), catalase (CAT), glutathione-S-transferase (GST), and lipid peroxidation in erythrocytes were determined in both seropositive and seronegative goats. Overall seroprevalence of O. ovis infection in goats was 56%. Positive significant (P<0.05) associations were observed among systemic IgG level and protein (0.34), hemoglobin (0.43), SOD (0.32), and MPO (0.41) in erythrocytes. Protein and hemoglobin concentrations, as well as SOD and MPO activities in erythrocytes were found significantly higher (P<0.05) in seropositive than in seronegative goats. By contrast, enzymatic activities of CAT and GST and lipid peroxidation values were similar in seropositive and seronegative groups. In conclusion, there was a systemic stimulation of Reactive Oxygen Species which was efficiently scavenged by erythrocytic antioxidant enzymes in goats seropositive to O. ovis infection.

Antibody responses to Lucilia cuprina in sheep selected for resistance or susceptibility to L. cuprina.

Sheep bred for resistance (R) or susceptibility (S) to fleece rot and myiasis (blowfly strike) have been shown to differ in inflammatory response to intradermal administration of blowfly (Lucilia cuprina) antigens and artificial challenge. The current paper describes analysis of antibody responses to L. cuprina antigens in the R and S animals. Serum antibody titres and specificities to larval antigens were examined and the specificity of wound exudate antibodies was also investigated in animals artificially challenged with L. cuprina. Titres of L. cuprina specific serum IgA, IgM, IgG2 and IgG1 were measured by ELISA, while specificities were examined on two-dimensional immunoblots of larval homogenates. Exposure to L. cuprina stimulated the production of specific antibody in both R and S animals, however antibody titres did not differ between the R and S animals. There was large variation in antibody specificity between individual animals and some L. cuprina proteins appear to be more frequently recognized by sera from either resistant or susceptible animals, however the recognition of a specific protein could not be solely attributed to the resistance status of the animal. It appears that resistance in these animals may be independent of serum antibody and is likely to be an innate response. Despite high levels of IgG in wound exudates, this antibody recognized few antigens in comparison with serum from the same animal, suggesting that exudate contains little functional antibody in comparison to serum.

Effect of myiasis and acute restraint stress on plasma levels of immunoreactive beta-endorphin, adrenocorticotrophin (ACTH) and cortisol in the sheep.

Cutaneous myiasis in sheep arising from the activity of Lucilia cuprina larvae can result in significant physiological changes in susceptible animals. The stress imposed on the pituitary-adrenal axis of the sheep in response to myiasis and acute restraint is the subject of this investigation. Merino wethers were exposed to handling restraint, and blood sampling, during examination for blowfly strike; where necessary, they were treated for cutaneous myiasis. Significant changes in the plasma concentrations of immunoreactive beta-endorphin (beta-EP), ACTH and cortisol were found in sheep with extensive myiasis, as compared with unstruck sheep or those with only localized myiasis. In five susceptible sheep with extensive cutaneous myiasis, mean plasma levels of beta-EP, ACTH and cortisol were 307 +/- 71 pg ml-1, 953 +/- 58 pg ml-1 and 232 +/- 46 nmol l-1 respectively, compared with 818 +/- 89 pg ml-1, 641 +/- 41 pg ml-1 and 107 +/- 17 nmol l-1 in six unstruck sheep handled similarly. Whereas significant increases in plasma ACTH and cortisol can result from pituitary-adrenal responses to acute emotional or surgical stress, and are usually accompanied by a concomitant release of beta-EP from the pituitary, the present findings indicate a marked reduction in beta-EP levels and a significant increase in ACTH and cortisol in sheep following blowfly strike and acute handling restraint. This result suggests that cutaneous myiasis in susceptible sheep can alter the pituitary-adrenal response to acute restraint stress, and this could occur either by an alteration of precursor processing in the pituitary or by the selective release of ACTH.

Expression of circulating leucocytes before, during and after myiasis by Dermatobia hominis in experimentally infected rats

Expression of circulating white blood cells was investigated in rats (Rattus norvegicus) experimentally infected with larvae of Dermatobia hominis, the human bot fly. Leucocytes were counted prior to infection (control group) as well as at 6, 10, 15, 20 and 28 days post-infection (dpi) and at 7, 15, 30 and 60 days post-larval emergence (dple). Total leucocyte numbers did not differ markedly among the groups. Significant differences were registered when values from control and animals harboring each larval stage of D. hominis were compared; with crescent rank: L1-, L2-, control and L3-infected groups. Leucocyte numbers were significantly higher in the control, 15, 20 or 28 dpi groups than in the 6 dpi animals. Higher counts were observed in control, L2- or L3-infected rats than L1-infected animals. Neutrophils, eosinophils and both large and small lymphocytes were also counted and analyzed. Basophils and monocytes were insufficient in number to permit statistical studies. These results stimulate the continuity of the studies about the host-parasite relationship in the dermatobiosis.

A expressão de leucócitos sangüínea foi investigada em ratos (Rattus norvegicus) experimentalmente infectados com larvas de Dermatobia hominis. As células foram contadas antes, durante, aos 6, 10, 15, 20 e 28 dias pós-infestação (dpi), e aos 7, 15, 30 e 60 dias pós-emergência das larvas dos hospedeiros. O total de leucócitos não apresentou marcante diferença entre todos os grupos de animais. Todavia, diferenças significantes foram observadas quanto ao parasitismo pelos estádios larvares, com nível crescente: L1, L2, controle e L3. Na comparação entre grupos: o número de leucócitos foi significativo pró-controle, -15, -20 ou -28 dpi do que aos 6 dpi; e pró-controle, -L2 ou -L3 do que para L1. Neutrófilos, eosinófilos e linfócitos (pequenos e grandes) foram também analisados. Em contraste, o número insuficiente de basófilos e monócitos não permitiram estudos estatísticos. Estes resultados estimulam a continuação dos estudos sobre a relação parasito-hospedeiro nas dermatobioses.