Intestinal mast cell response in thymectomised and normal mice infected with Trichinella spiralis.

In NIH mice, expulsion of $\text{Trichinella spiralis}$ from the small intestine and increase in intestinal mast cells were each dependent on the presence of T-lymphocytes. Both changes were deficient in thymectomised mice but could be largely restored by reconstitution of thymectomised mice with syngeneic mesenteric lymph node cells. In both NIH and BALB/C mice the majority of the increased number of mast cells occurred within the intestinal epithelium. In NIH mice increase in the number of intestinal mast cells coincided roughly with expulsion of the parasites.In BALB/C mice increase in the numbers of intestinal mast cells did not appear to be connected with the location or expulsion of parasites and it is concluded that mast cell proliferation, accumulation and discharge $\text{per se}$ do not result in or from worm expulsion.     

Coccidiosis: T-lymphocyte-dependent effects of infection with Eimeria nieschulzi in rats

The intestinal pathology caused by infection with $\text{Eimeria nieschulzi}$ was investigated and comparisons were made between the effects in athymic nude (rnu/rnu) rats and their heterozygous (rnu/+) litter-mates. Most of the changes noted, i.e. increase in gut weight, partial villous atrophy and increased numbers of mast, goblet and pyroninophilic cells were shown to be largely or wholely thymus dependent. The numbers of intraepithelial lymphocytes were decreased in both groups during the period of study. The peripheral blood leucocyte response was similar in both groups of rats during a primary infection but differed after a challenge inoculum, indicating that the secondary type of response which occurred in the rnu/+ rats was thymus dependent, as is resistance to reinfection.     

Nippostrongylus brasiliensis infection in the rat: 2. A comparison of production of antibodies in bile and serum by baby rats,lactating rats and adult rats

Production of bile lgA and serum lgG antibody responses against whole worm antigens and worm metabolic product antigens occurred by 7 to 14 days after a single infection with $\text{N. brasiliensis}$ and was similar in adult rats, lactating rats and baby rats. Production of bile lgA antibodies against phosphorylcholine was greater in adult rats, 7 days after infection, than in baby rats or lactating rats. lgA antibody production by baby rats may be affected by immaturity of the intestinal lgA system, and diversion to the mammary gland of lgA-plasma cell precursors may occur in lactating rats. Such effects may be involved in the failure of baby rats and lactating rats to expel parasites by day 21 at which time in adult rats expulsion was complete.     

Nippostrongylus brasiliensis infection in the rat: 1. Production of bile lgA and serum lgG antibodies by adult rats

Bile lgA and serum lgG antibody responses were assayed by a micro ELISA technique. Antibodies of both classes were produced by 14 days after infection against whole worm antigens and worm metabolic product antigens, by adult rats infected once with $\text{Nippostrongylus brasiliensis}$. Rats were reinfected on day 28 and titres of both classes of antibody against both antigens were greater 7 and 14 days after reinfection than after the initial infection. Bile lgA antibodies were produced against phosphorylcholine after a single infection with the parasite but no increased response occurred after reinfection. The possible significance of lgA and lgG antibody responses in relation to immunity to intestinal nematode infections is discussed.     

Lack of reactivity of sera from Theileriaparva-infected and recovered cattle against cell membrane antigens of Theileria parva transformed cell lines

Sera from $\text{Theileria}$$\text{parva}$ infected, recovered and rechallenged cattle were tested in complement-dependent cytotoxicity, membrane immunofluorescence and antibody-dependent cellular cytotoxicity assays for the presence of antibodies against cell membrane antigens of $\text{T}$. $\text{parva}$ transformed cell lines.In the complement-dependent antibody-mediated cytotoxicity assay, sera from lethally infected animals were negative. Some recovered cattle showed a positive reaction, but such reactions were also observed when an eland cell line infected with $\text{T}$. $\text{taurotragi}$, and bovine lymphoblastoid cells were used as targets. Reaction was less against Ig-negative peripheral blood lymphocytes.Evidence is presented that these reactions could be evoked by attachment of immune complexes to Fc-receptors. It is concluded that cattle exposed to $\text{T}$. $\text{parva}$ infection do not develop antibodies against specific $\text{T}$. $\text{parva}$ (or $\text{T}$. $\text{parva}$-induced) cell surface antigens.     

The influence of concurrent infection with Oesophagostomum species on the interpretation of the Elisa test for trichinosis in pigs

The effects of simulatenous infection of pigs with Oesophagostomum spp. and Trichinella spiralis on the interpretation of the Enzyme-Linked Immunosorbent Assay (ELISA) for trichinosis were examined. Extinction values were observed from four groups of pigs. The first group acted as uninfected controls, the second was infected only with Oesophagostomum spp., the third with T. spiralis alone and the fourth by both nematodes.It was found that the pigs infected with T. spiralis could be differentiated from the others, but that those infected with both species had lower extinction values than the group with T. spiralis alone. The differences may be related to the numbers of T. spiralis larvae able to establish and develop into adults in the small intestine of the host. Those infected with Oesophagostomum spp. alone showed no rise in extinction values, and it was concluded that there was no cross-reaction in the ELISA between thos pecies and T. spiralis antigen.     

The use of a cathodic antigen in the immunoelectrophoretic serodiagnosis of Echinococcus granulosus in sheep

An antigen was prepared from purified sheep hydatid-cyst fluid by gelfiltration on Sephadex G-200 followed by chromatography on DEAE-cellulose. In each case the first-peak material was used. This antigen, which migrated cathodically, was concentrated and used in immunoelectrophoretic analyses of 4X concentrated sera from sheep experimentally and naturally infected with $\text{Echinococcus granulosus}$ and from uninfected sheep. Of 34 sheep with $\text{E. granulosus}$ infection 31 were positive with the cathodic antigen while of 85 sheep without $\text{E. granulosus}$ 8 were (falsely) positive. Many false positives appeared to be associated with heavy infections of $\text{Taenia ovis}$ or $\text{T. hydatigena}$ larvae. The “arc 5” immunoelectrophoresis test, which is the most specific immuno-diagnostic test for echinococcus infection in humans, was not able to specifically identify $\text{E. granulosus}$ infections in sheep.     

Bovine babesiosis: The immunization of cattle with fractions of erythrocytes infected with Babesia bovis (syn B. argentina)

Soluble antigen which protected susceptible cattle against challenge with $\text{Babesia bovis}$ was extracted from $\text{B. bovis}$-infected erythrocytes by sonic disintegration and separation of the soluble from the insoluble matter by ultracentrifugation. The material was then fractionated by the precipitation of fibrinogen-like proteins. The precipitate contained the babesial antigens that were located on the stroma of the infected erythrocytes. Antigen originally located on the parasite remained in solution. Both fractions conferred protection on splenectomized calves against challenge with $\text{B. bovis}$. However, the fraction containing the parasite antigens appeared to have more potential for development as a killed vaccine because it was not heavily contaminated with antigenic material from bovine erythrocytes.     

Application of indomethacin as a potentiator of lymphocyte blastogenesis in Brucella abortus exposed cattle

Lymphocytes from $\text{Brucella abortus}$ field strain infected, strain 19 vaccinated, non-exposed and field strain infected, but immunologically unresponsive cattle were incubated with $\text{B. abortus}$ antigen and indomethacin. There were significant increases (P < 0.005) in the blastogenic responses, as measured by [³H] thymidine uptake, in cultures with indomethacin as compared to cultures without indomethacin. Lymphocyte blastogenic responses to $\text{B. abortus}$ antigen were potentiated by indomethacin in both $\text{B. abortus}$ exposed and non-exposed cultures. However, potentiation of sensitized lymphocyte blastogenic responses by indomethacin was significantly greater (P < 0.005) than that in non-exposed lymphocytes. Additionally, indomethacin significantly potentiated Brucella-induced lymphocyte blastogenic responses in lymphocytes from anergic cattle.     

Antigens and antibodies of malignant catarrhal fever herpesvirus detected by immunodiffusion and counter-immunoelectrophoresis

Using hyperimmune rabbit and cattle sera, immunodiffusion (ID) and counter-immunoelectrophoresis (CIEP) tests detected three or four and two or three malignant catarrhal fever (MCF) virus antigens, respectively, in infected cells. The ID test detected precipitating antibodies to MCF virus in $\text{3}\text{9}$ experimentally infected rabbits, $\text{0}\text{14}$ experimentally infected cattle, $\text{1}\text{13}$ naturally infected cattle, $\text{62}\text{176}$ wildebeest and $\text{3}\text{20}$ hartebeest. The CIEP test detected specific antibodies in $\text{3}\text{9}$ rabbit sera, but non-specific reactions prevented its use with bovine sera.The CIEP test was 2 to 4 times more sensitive than ID for detecting antibodies to MCF virus, but both tests were less sensitive than indirect immunofluorescence.The ID test demonstrated an antigenic relationship between wildebeest and hartebeest strains of MCF virus. Neither ID nor CIEP detected MCFV antigens in tissues infected with MCF virus.     

Effect of phytohaemagglutinin-P on apoptosis and necrosis in Trichinella spiralis infected mice

Flow cytometry analyses were used to evaluate the contribution of apoptotic and necrotic lymphocytes in the selected organs of Trichinella spiralis infected mice treated with phytohaemagglutinin-P (PHA-P). The Tunnel method was used to examine apoptosis in a cryostat section from the jejunum and masseter muscle. CFW mice (Groups I and II) were infected with 200 larvae of T. spiralis. PHA-P was administered intravenously at a dose of 10 mg/kg 24 h prior to infection in Group II mice only. Group III mice were treated with PHA-P without T. spiralis infection, and Group IV mice were untreated controls. The lymphocytes obtained from the spleen, mesenteric lymph nodes (MLN) and muscular inflammatory infiltration on 7, 14, 21, 28, 35, 42 and 60 days post infection (DPI) were incubated with the Annexin-V-Fluos Staining Kit (Roche). The cryostat preparation made from the jejunum and masseter muscle was evaluated using a fluorescence microscope. PHA-P administration stimulated apoptosis in the jejunal mucosa and in the muscular inflammatory infiltration. In Group I mice, infected with T. spiralis only, the highest percentage of apoptotic cells was found on 7 DPI in the spleen and in MLN, and on 14 DPI among the cells of the muscular inflammatory infiltration. The peak of the necrotic lymphocytes was found on 7 DPI in the spleen, on 28 DPI in MLN, and on 21 DPI in the cells of muscular inflammatory infiltration. In Group II mice, infected with T. spiralis and treated with PHA-P, the peak in apoptotic cells occurred on 7 DPI in the spleen and in the muscular inflammatory infiltration. The highest level of necrotic lymphocytes was observed only on 7 DPI in the muscular inflammatory infiltration. Percentage of necrotic lymphocytes in the spleen was the same and in MLN it was lower than in Group I (T. spiralis only). Moreover, the number of muscle larvae in mice treated with PHA-P (Group II) was lower than in Group I (T. spiralis only).     

Eosinophil relationship in gut anaphylaxis during experimental ancylostomosis

Changes induced in the circulating and intestinal eosinophil levels and worm load were studied in female Swiss albino mice infected with multiple doses of Ancylostoma caninum larvae. There was a significant rise of both circulating and intestinal eosinophils in all the infected animals when compared with controls. In all the experimental groups of animals, peak eosinophilia was observed during the third week of post-infection and sustained upto 30th day with slight alterations. These studies indicated that eosinophilia will be induced due to anaphylactic reactions in the gut and eosinophilia in multiple infections is an anamnestic response.     

Brucella abortus antigen-induced lymphocyte reactivity in guinea pigs

A whole blood lymphyocyte transformation (WBLT) assay was used to detect anti-brucella lymphocyte reactivity in guinea pigs. Brucella antigens stimulated an antigen-specific lymphoproliferative response in WBLT assays from $\text{Brucella abortus}$ infected guinea pigs. The response was best detected from 6 to 16 weeks after challenge inoculation with viable $\text{B. abortus}$ 2308. Lymphocytes were not stimulated by unrelated bacterial antigens and control animals did not respond to the Brucella antigens. The responding cell population was characterized as mostly T lymphocytes. The WBLT assay was found to be specific for the detection of anti-brucella lymphocyte reactivity. However, a negative response was not definitive, which indicated a need for repeated testing to establish that a guinea pig did not have anti-brucella lymphocyte sensitization.     

Trichuris suis population dynamics following a primary experimental infection

Trichuris suis population kinetics was studied by experimentally infecting 40 pigs with 5000 T. suis infective eggs. Six pigs were sacrificed every 2 weeks from 1 to 9 weeks post-inoculation (p.i.) and the remaining 10 pigs were sacrificed 11 weeks p.i. to estimate worm burdens. An equal number of uninfected control pigs were sacrificed at the same time points for comparison. Egg excretions from each pig were evaluated every week from 5 to 11 weeks p.i. Peripheral blood eosinophilia and basophilia were also evaluated every 2 weeks throughout the experimental period. After an initial almost 100% establishment of T. suis an expulsion phase followed approximately 9 weeks p.i., resulting in an aggregated distribution of the worms in the pig population, as it is characteristic for most helminth infections. By 11 weeks p.i. almost all worms had been expelled. Egg excretion peaked 7 weeks p.i. and a significant non-linear relationship between worm burdens and egg excretion was found. The predilection site for T. suis was the caecum and proximal colon and the relative distribution of worms in the large intestine did not change over time until expulsion. Peaking peripheral blood eosinophil and basophil levels were observed in the T. suis infected pigs 5 and 7 weeks p.i., respectively, reflecting the host activated immune response against the parasite. We here describe the course of a primary T. suis infection in pigs by detailed worm counts, demonstrating an effective expulsion that results in an almost 100% clearance of infection as previously indicated by monitoring faecal egg excretion.     

Experimental production of congenital persistent swine fever infections: I. Clinical,pathological and virological observations

Twelve sows were inoculated with a low-virulent field strain of swine fever (SF) virus at 40, 65 and 90 days of pregnancy. Transplacental infection occurred in eight sows and these farrowed 88 piglets.Prenatal mortality was highest in litters from sows infected at 40 days after service, and postnatal death was most frequent in litters from sows infected at 65 days. Three sows gave birth to completely infected litters, whereas the five others farrowed litters in which uninfected piglets were found. The later the sows were infected during pregnancy, the more uninfected piglets were born. Twelve piglets recovered from the infection and the percentage of recoveries increased with the stage of pregnancy at which inoculation took place. Twenty-three piglets developed a persistent infection. The earlier infection occurred during pregnancy, the more persistent infections were produced. The persistently infected pigs developed a runting syndrome from about one week after weaning. The clinical signs were more severe in pigs from sows infected at 65 days of pregnancy than in pigs from sows infected at 40 days. A persistent viraemia was present in these pigs with titres ranging between 10^5.7 and 10^7.0 plaque forming units/ml of plasma. At autopsy the most pronounced lesions were an atrophy of the thymus, and swollen and pale lymph nodes. Virus antigen was present in lymphoidal, reticulo-endothelial and epithelial tissues.One persistently infected pig survived superinfection with a virulent strain of SF virus for 45 days.     

The effect of γ-radiation on the development of infective larvae of Trichostrongylus colubriformis in guinea pigs and sheep

Infection of guinea pigs with normal Trichostrongylus colubriformis larvae was characterized by an initial logarithmic decline of total worms recovered over the first few days of infection (“loss phase”) followed by a “plateau phase” when worm numbers remained stationary and a third “expulsion phase” when worm numbers again declined logarithmically with time. Irradiation of infective larvae at 50 and 100 kilorads (krad) affected the subsequent pattern of infection by prolonging the initial “loss phase” and contracting the “plateau phase”. The rate of decline of total worms during either the “loss phase” or the “expulsion phase” was not affected by larval irradiation.Few males were recovered at any stage of infection with larvae irradiated at 50 or 100 krad. However, the irradiated larvae (females) which survived continued development at a rate which was similar to that of normal larvae and it was apparent that these parasites were not able to maintain their position in the small intestine, but continued development during migration towards the posterior.The radiosensitivity of male larvae and worm egg production (eggs in utero) over a range of radiation doses (25 to 100 krad) were remarkably similar. On the other hand, female larvae were clearly less sensitive to γ-radiation. These observations suggested that the depressed egg production in females may be due to the absence of males.     

Quantitative studies of ovine haemonchosis. I. Relationship between faecal egg counts and total worm counts

The successful operation of parasite-control programmes based on monitoring of faecal egg counts stimulated an examination of the quantitative relationship between egg counts and worm counts of Haemonchus contortus in sheep in the Goondiwindi area of south-east Queensland. Sixty-one commercial Merino sheep were selected from flocks where outbreaks of haemonchosis had occurred. Faecal samples were collected prior to euthanasia and eggs and worms subsequently counted and differentiated.A strong relationship ($\text{r}{}^2\text{= 0.83, 71P < 0.001}$) was found between faecal egg counts and the total number of adult H. contortus. The size of the worm population, the time of year, and the ratio of male to female adult worms were minor sources of variation. The relationship was considered to be of value in the implementation of programmes for the diagnosis and control of haemonchosis in sheep flocks in the area.     

Murine model study of the practical implication of trypanosome-induced immunosuppression in vaccine-based disease control programmes

The relevance of trypanosome-induced immunosuppression in relation to the efficacy of vaccine-induced immunity was studied in mice. Mice were immunised with crude Trichinella spiralis muscle larvae homogenate vaccine and infected with T. spiralis and/or Trypanosoma brucei. Vaccination significantly decreased adult worm burden (p<0. 05) and accelerated worm expulsion in mice infected with T. spiralis only. T. brucei superinfection resulted in monocytosis, suppressed eosinophilia, significant decrease in PCV (p<0.001), higher numbers of adult worms (p<0.001) and failure to expel all adult worms by Day 12 post infection (p.i.). Regardless, they produced anti-Trichinella IgG(1) responses similar to those of the vaccinated non-T. brucei-infected group. T. brucei also suppressed the proliferative responses of spleen cells to stimulation with Con A and T. spiralis antigen, and induced strong production of interferon-gamma (IFN-gamma) in culture supernatants of antigen stimulated spleen and mesenteric lymph node cells. Interleukin-5 (IL-5) production was suppressed by T. brucei in supernatants of Con A- and antigen-stimulated spleen cells. It was concluded that trypanosome infections and the associated immunosuppression are of great practical significance in trypanosome endemic areas, especially with regards to disease control programmes involving vaccine-induced herd immunity.     

Solid-phase enzyme immunoassay of bovine antibody responses following immunization against and natural infection with Pasteurella haemolytica

A solid-phase enzyme immunoassay (EIA) was developed in order to monitor bovine antibody responses following immunization against and natural infection with $\text{Pasteurella haemolytica}$ serotype A:1. Non-ionic surfactants, used in many antibody EIAs to reduce non-specific immunoglobulin binding, had to be avoided because they inhibited specific binding of bovine antibodies to $\text{P. haemolytica}$ antigens. Calves were immunized with a KSCN extract of $\text{P. haemolytica}$. Subcutaneously immunized animals developed a significantly higher humoral antibody response than did intranasally vaccinated animals. Intranasally immunized calves developed a slightly, but not significantly higher nasal antibody response than did calves vaccinated subcutaneously. Field study results based on bacterial isolation and EIA detection of antibodies to $\text{P. haemolytica}$ indicate that cattle can generate carrier states where bacteria are present in the upper respiratory tract, yet no humoral antibody response is induced. The converse was also found where cattle were free from $\text{P. haemolytica}$ in the upper respiratory tract, yet possessed a good humoral antibody response to $\text{P. haemolytica}$.