The elimination of equine strongyles and hematological and pathological consequences following larvicidal doses of thiabendazole

Twelve horses wer divided into three groups and given various doses of a mixed species strongyle inoculum, representing light, moderate, and heavy infctions. Three weeks after the larval inoculations, three animals from each group were given larvicidal doses of thiabendazole (TBZ) (440 mg kg^?1 on two consecutive days); one animal from each group served as a non-medicated control. Treatment was repeated three weeks later. One treated animal from each group was designated for long-term study; others were necropsied to study adult and larval parasite loads.Six of the twelve animals with strongylosis developed moderate eosinophilia.TBZ given at 440 mg kg^? on two consecutive days caused depression, lethargy, and anorexia which lasted for five days. Eosinopenia, lymphopenia, and neutrophilia occured in treated animals, and lasted for three days. During the course of TBZ treatment, one horse died from what appeared to be a mis-dosing or an anaphylactic reaction.At necropsy, active thrombi of the anterior mesenteric artery were seen in parasitized animals, but not in those treated with TBZ. Five out of seven medicated horses were completely free of adult and larval strongyle parasites. One had a few STrongylus edentatus larvae and another had small strongyles. No Strongylus vulgaris larvae or adults were recovered from any horse treated with TBZ.     

The role of lipoteichoic acids on the adherence of Streptococcus equi to epithelial cells

The effect of the presence of lipoteichoic acids (LTA) on a strain of Streptococcus equi was investigated. The LTA were extracted in a crude form from the S. equi strain and were found to sensitize sheep red blood cells so that they agglutinated with antibodies specific to purified LTA of group A streptococci. The crude LTA preparation was also able to inhibit the specific haemagglutination reaction involving group A streptococcal LTA and LTA antibodies. Neither the purified LTA from group A streptococci nor the anti-LTA serum interfered with the adherence of S. equi to equine epithelial cells.     

The migration of larval Toxocara canis in mice II. Post-intestinal migration in primary infections

Following oral infection of NIH mice with Toxocara canis embryonated eggs the L₂ pass the visceral phase of migration during the first week of infection. Larvae reach the liver and lungs and peak in number in these organs 2 and 3 days after infection, respectively. Larvae are then dispersed throughout the body and enter the myotropic—neurotropic phase by the 7th day of infection. Larvae injected directly into the brain are capable of migrating into the viscera and musculature. Considerable pathology occurs due to larval migrations, especially through the liver and lungs, and both acute and chronic disease are recorded. Studies of infections extending over a year show that the number of recoverable larvae declines gradually with periods of stable populations.On Days 3, 4 and 5 after infection, larvae were demonstrable in the faeces of infected mice. Prenatal infection was observed in a third of the offspring of mice infected the same day as conception.     

The effects of dl-tetramisole and rafoxanide on tricarboxylic acid cycle enzymes of Haemonchus contortus, in vitro

Various enzymes of the tricarboxylic acid cycle (TCA) viz., aconitase (E.C. 4.2.1.3), isocitrate dehydrogenase (E.C. 1.1.1.42), succinate dehydrognease (E.C. 1.3.99.1), fumarate reductase (NADH: fumarate oxido-reductase), fumarase (E.C. 4.2.1.2) and maltate dehydrogenase (E.C. 1.1.1.37) were detected in adult Haemonchus contortus (Nematoda: Trichostrongylidae), in vitro. Low activities of aconitase and isocitrate dehydrogenase suggested that the TCA cycle has a minor function and the pathway of CO₂ fixation is the major pathway in the energy metabolism of the parasite. In vitro incubation in Tyrode's solution had no significant effect on TCA cycle enzymes and the worm was able to maintain normal metabolism for 12 h.The effects of dl-tetramisole and rafoxanide on various enzymes of the TCA cycle were studied in adult H. contortus. At 50 μg ml^?1 varying degrees of succinate dehydrogenase and fumarate reductase activities were observed. At the same concentration, the activities of other enzymes remained unaltered.     

Adherence of Streptococcus equi on tongue,cheek and nasal epithelial cells of ponies

Streptococcus equi was found to adhere to tongue, cheek and nasal epithelial cells of ponies, in vitro. Maximum adherence was observed at pH 7.5 after one hour of incubation of bacteria with epithelial cells. This adherence was more on epithelial cells from adult animals than foals. Streptococci exposed to heat (60°C for 10 min) or treated with pepsin or trypsin showed a reduced adherence, whereas an increase occurred on treatment with hyaluronidase. Antibodies against whole S. equi cells or M-like protein blocked the adherence, whereas antibodies against group-specific carbohydrate or lipoteichoic acids did not. Pretreatment of epithelial cells with either the M-like protein or crude extract of S. equi lowered the adherence, whereas an extract of S. zooepidemicus did not. Adherence of S. equi to the epithelial cells was considered to be mediated by structures specific to S. equi.     

A serological and biochemical study of new field isolates of foot-and-mouth disease virus type A in Peru, 1975 to 1981

Three foot-and-mouth disease virus type A isolates recovered from field outbreaks in the Department of San Martin, Peru, during the period 1975 to 1981 were compared with each other, and the South American vaccine strains A24 and A27, by complement fixation (CF), virus neutralization (VN) and polyacrylamide gel electrophoresis (PAGE). Complement fixation and VN tests gave comparable results distinguishing the field isolates from each other and from the vaccine strains. Analysis of the structural polypeptides by PAGE also showed clear differences between all the viruses examined. Samples from tissue culture passaged and mouse adapted strains of one of the field isolates gave identical patterns in PAGE, but differences were observed in the polypeptide pattern of the A24/BRA/55 strain and the Peru vaccine strain, which were serologically indistinguishable. Results illustrate a continued antigenic variation in an endemic area where vaccination has been used; however, asymmetric serological reactions between the A24 vaccine strain and the most recent field isolate indicated that a vaccine incorporating A24 should still give adequate protection.     

Experimental and natural infection of calves with Bunostomum phlebotomum

An experiment was conducted to determine the efficiency of a method of experimental infection of weaner beef calves with Bunostomum phlebotomum and to compare such infection with that established by natural infection. Six calves, maintained on a concrete-floored pen, were inoculated with B. phlebotomum L3 by placing the larval inocuulum, in small volume, in the outer chamber of the ear while the animal was restrained for 18 min. Inoculation doses of 20, 30, 40, 50, 60, and 80 thousand L3 were used. Six other calves were grazed on pasture known to be heavily contaminated with hookworm. All animals were killed 72 days after experimental infection and 93 days after initial exposure to pasture infection. The experimental and naturally-infected calves became patent at 55 and 66 days, respectively, after exposure to L3. Red blood cell counts and hemoglobin values were markedly depressed in both groups and lowest values coincided with onset of patency. There was difference in liveweight changes, but both groups lost weight during the prepatent period of infection and gained weight with the onset of patency. The largest number of hookworms was established at the 30000 L3 inoculation level; little or no establishment was observed at the 2 highest levels. Sizeable adult hookworm burdens were established in 4 out of 6 pastured calves. Intestinal pathology was generally more severe in experimentally-infected calves, consisting of a thickened mucosa and masses of punctate, hemorrhagic foci. Pastured calves also acquired large burdens of Ostertagia ostertagi, particularly inhibited early fourth-stage larvae. Moderate to severe abomasal pathology and elevated plasma pepsinogen were associated with ostertagiasis in the pastured calves. The experimental infection method is efficient in establishing high levels of B. phlebotomum infection in calves currently or previously infected with other gastrointestinal nematodes.     

Morphogenesis of Trichostrongylus rugatus and distribution during development in sheep

Morphogenesis of Trichostrongylus rugatus was examined in 16 sheep experimentally-infected with 120 000 third-stage larvae and killed 2, 4, 6, 8, 10, 12, 16 and 20 days later (DAI). Third stage larvae moulted between Days 4 and 6, and fourth stage larvae moulted on Day 10 after infection. Four sheep first passed eggs in the faeces between Days 16 and 18 after infection. Rate of growth of larvae was constant between 2 and 10 DAI followed by a period of rapid growth from 10 to 16 DAI. Major features of larval development are described. Nematodes were largely restricted to the first 6 m of gut with 71% of worms occurring in the first 3 m.     

Ecology of Rhodococcus equi

A selective broth enrichment technique was used to study the distribution of Rhodococcus equi in soil and grazing animals. Rhodococcus equi was isolated from 54% of soils examined and from the gut contents, rectal faeces and dung of all grazing herbivorous species examined. Rhodococcus equi was not isolated from the faeces or dung of penned animals which did not have access to grazing. The isolation rate from dung was much higher than from other samples and this was found to be due to the ability of R. equi to multiply more readily in dung. Delayed hypersensitivity tests were carried out on horses, sheep and cattle, but only horses reacted significantly. The physiological characteristics of R. equi and the nature of its distribution in the environment suggested that R. equi is a soil organism.     

Fine structure and invasive behaviour of the early developmental stages of Theileria annulata in vitro

The interaction, in vitro, between bovine peripheral blood lymphocytes and sporozoites of Theileria annulata (Ankara) was studied by light and electron microscopy. Beginning five minutes following incubation, samples were taken for Giemsa-stained smears and glutaraldehyde-fixed pellets, for light and electron microscopy, respectively. Sporozoites of T. annulata measure an average of 0.9 microns long, 0.8 microns broad and possess a limiting unit membrane, the pellicle; a round-to-ovoid, eccentrically situated, non-chromocentric nucleus; double-membraned, tubular, acristate mitochondria; varying numbers of anisocytic, densely osmiophilic and pleomorphic organelles, the rhoptries which together with the polar ring form the apical complex; and numerous, loosely scattered, electron-dense ribosomal particles. As early as 5 min of incubation, sporozoites had made contact with, and penetrated, lymphocytes. Sporozoites consistently attached to the lymphocyte plasmalemma by their basal end, possibly at specific receptor sites. Apparently only a proportion of lymphocytes (up to 40% and more commonly 10-20%) were susceptible. Two subpopulations of the susceptible lymphocytes were observed; one which appeared to have receptor sites localized on one pole of the plasmalemma and the other subpopulation in which the receptor sites were distributed evenly around the plasmalemmal surface. Within individual susceptible lymphocytes, the number of interiorized sporozoites increased from 1 to 3 at 5-10 min to as many as 15 or more parasites at around 60 min of incubation. Theileria annulata sporozoites were interiorized by the invagination of the host cell plasmalemma which remained intact throughout the process but later fragmented. Within 30 min of interiorization, each sporozoite underwent dedifferentiation by the loss of its rhoptries and transformed into a trophozoite. Around 24 h, the trophozoite, a uninucleate, motile and feeding stage of the parasite, developed into a schizont by an acentric, closed mitosis.     

Isolation of Fasciola hepatica metacercariae by density gradients

Fasciola hepatica metacercariae were purified in high yield, removing contaminating cyst walls and plant material by step gradients consisting of 10 ml of 60% Percoll (density = 1.08 g ml-1) and 10 ml of 50% Metrizamide (density = 1.25 g ml-1). Greater than 90% of the metacercariae applied to the density gradients were recovered. These isolated metacercariae had an in vitro excystment rate of greater than 80%, which was the same excystment rate as metacercariae not subjected to density gradient centrifugation.     

Experimental infection of pups with Ancylostoma caninum-infected mouse tissues

Migration and distribution of Ancylostoma caninum larvae in the tissues of mice orally infected with 1000 larvae, and establishment of patent infection from the mice to the definitive host, were studied. Larval yield from different organs of mice, after digestion with artificial gastric juice, indicated that the highest recovery was at 4 h post-infection (62.8%), and thereafter a slight decline occurred up until 30 days post-infection (51.5%). Migration of larvae to the lungs occurred within 4 h, to the liver within 12 h and into the heart within 24 h. No larvae were recovered from spleen and kidney tissues. From the 9th day onwards larvae were also recovered from the brain. Migration in the muscles of head and neck occurred within 4 h, in the thoracic and abdominal muscles at 24 h and in lumbosacral and leg muscles at 48 h. The establishment of patent infections in the definitive host was studied by feeding the orally- and percutaneously-infected mice to hookworm-free pups at 10 and 30 days post-infection. The mean necropsy worm burden in the pups fed with the orally-infected mice was comparatively higher than in the pups fed cutaneously-infected mice.     

Localization of culture-derived soluble Babesia bovis antigens in the infected erythrocyte

Immunoprecipitates derived from crossed immunoelectrophoresis of Babesia bovis culture supernatant fluid against a polyspecific anti-B. bovis serum were used to produce monospecific rabbit antibodies to individual B. bovis antigens. These antibodies were utilized in an immunofluorescence test to identify the location of the respective antigens within the infected erythrocyte. Two antigens were found on or near the erythrocyte membrane, while a third antigen was directly associated with the parasite itself.     

Serum and abomasal antibody response of sheep to infections with Haemonchus contortus

Serum and abomasal IgA, IgG and IgM antibody response against adult worm, L3 and egg antigens of Haemonchus contortus was monitored by the ELISA technique after one or two infections with this nematode. Following the first infection, antibody levels in serum did not change materially. After administration of a challenge dose of infective larvae, antibodies of the three immunoglobulin classes in infected animals rose slightly, but this rise appeared later than the fall in the faecal egg counts. In contrast, in abomasal mucosa, IgA anti-larval antibody levels, which did not increase materially after the primary infection, rose rapidly after a transient inhibition when sheep were challenged. A close temporal relationship was observed between the rise in local anti-worm IgA antibodies and the self-cure reaction, but antibody levels fell rapidly after worm diminution. The local antibody response was thus considered to be related to immunity of sheep to H. contortus.     

Survival and infectivity of Dicrocoelium dendriticum eggs under field conditions in NW Spain

A study was made of the survival of Dicrocoelium dendriticum eggs in sheep faeces in an area with a temperate climate (NW Spain). It appears that mortality is independent of the age of the eggs and that there is a marked seasonality within the period of time considered (20 months). A study was also carried out on the infectivity of D. dendriticum eggs by experimental infections of suitable intermediate hosts. The data show that no loss took place in infectivity during the period of study (15 months).     

Isolation of enterotoxigenic strains of staphylococci from dogs

The ability of 309 staphylococcal isolates from household dogs to produce enterotoxin, coagulase, thermonuclease and hemolysin was investigated. A total of 52 (16.8%) isolates from 45 out of 150 dogs examined were enterotoxigenic when tested for enterotoxin types A, B and C. Based on sites sampled, 33 (20.5%) out of 161 isolates from the anterior nares were enterotoxigenic while from dorsal skins 19 (12.8%) out of 148 isolates were enterotoxigenic. Staphylococcal enterotoxin C(SEC) was predominantly produced as 21 (6.8%) isolates elaborated it and also accounted for 40.4% of all enterotoxins produced by isolates. Staphylococcal enterotoxins A(SEA) and B(SEB) were produced by 10 (3.2%) and 16 (5.2%) strains, respectively. Mixed enterotoxin types AB, AC and BC were produced by 1,3 and 1 strains, respectively. With human plasma, 17.1% of coagulase-positive and 15.0% of coagulase-negative strains were enterotoxigenic. However, using canine plasma, 19.1% and 6.9% of the coagulase-positive and negative isolates, respectively, were enterotoxigenic. The incidence of enterotoxigenicity was 16.9% amongst thermonuclease-positive isolates and 16.3% for thermonuclease-negative strains.Alpha hemolysin was predominantly produced by 180 (60.2%) isolates and 19.9% of these were enterotoxigenic. Beta hemolysin was produced by 36 (11.7%) isolates with 13.9% enterotoxigenic, while 87 (28.2%) exhibited gamma hemolytic pattern amongst which 11.5% were enterotoxigenic.Based on data provided on coagulation of human and canine plasmas and hemolytic patterns, it is concluded that a large proportion of canine isolates from this environment are not of canine biotypes, but are most probably human biotypes.     

Pathogenicity of Australian isolates of Haemophilus paragallinarum and Haemophilus avium in chickens

Twenty-seven Australian avian Haemophilus isolates were tested for their ability to cause infectious coryza in specific pathogen-free chickens. All 15 isolates, identified as H. paragallinarum, produced infectious coryza, whereas all 12 H. avium isolates were nonpathogenic, but spread to in-contact chickens.     

Acquired immunity to Eimeria tenella in lasalocid-treated chickens

The effect of restricted medication with lasalocid sodium on the development of acquired immunity against Eimeria tenella was evaluated. The medication was allowed for all or part of 6-day test period (one day before until 4 days after infection). The parameters used for such evaluations were lesion score, caecal, bursal and splenic weights. The optimum treatment time for the drug was clearly indicated by lesion score which was very low when the medication was initiated 1 day before until 1 day after inoculation, but only partly effective if given on Day 2 post-inoculation. The challenge with higher doses on 14th day of immunizing infection revealed a reverse picture where the higher lesions were recorded by the groups where medication was started earlier than the delayed treatment groups. This indicates partial interference with the development of immunity in the earlier treatment groups. Birds treated on Day 4 p.i. were not significantly different (P less than 0.05) from the infected unmedicated control group, suggesting no interference in acquired immunity. A correlation was noticed between day of treatment, the lesion score and weight gain of the caecum as well as the spleen. After both immunizing and challenge infections, the bursa did not show any significant variation in weight, whereas the weight of the spleen did vary. The infected unmedicated group and the delayed-treatment groups had a comparatively higher splenic weight than the uninfected unmedicated group of birds.(ABSTRACT TRUNCATED AT 250 WORDS)     

T- and B-lymphodi cell population in calves immunized against Theileria annulata

Distribution of T- and B-lymphoid cells in peripheral blood, lymph node, spleen and bone marrow of normal healthy calves and calves immunized against Theileria annulata followed by challenge on Day 50 post-immunization were studied by rosette tests. Significantly increased percentages of T- and B-lymphoid cells were recorded in immunized calves.     

Non-specific immunization against bovine tropical theileriosis (Theileria annulata) using killed Corynebacterium parvum

Killed Corynebacterium parvum was used as an adjuvant for the production of non-specific resistance against Theileria annulata in cattle. Groups of cross-bred (Bos indicus X Bos taurus) calves were administered C. parvum adjuvant subcutaneously and were then challenged with T. annulata-infected ticks on 45, 60 or 90 days later. The challenge caused mild reactions in the protected calves. None of the 10 immunized calves died due to theileriosis, whereas all three paris of susceptible control calves died due to theileriosis. It appears from this pilot study that cattle can be protected non-specifically with C parvum adjuvant against T. annulata.