Evidence of endemic Hendra virus infection in flying-foxes (Pteropus conspicillatus)--implications for disease risk management

This study investigated the seroepidemiology of Hendra virus in a spectacled flying-fox (Pteropus conspicillatus) population in northern Australia, near the location of an equine and associated human Hendra virus infection in late 2004. The pattern of infection in the population was investigated using a serial cross-sectional serological study over a 25-month period, with blood sampled from 521 individuals over six sampling sessions. Antibody titres to the virus were determined by virus neutralisation test. In contrast to the expected episodic infection pattern, we observed that seroprevalence gradually increased over the two years suggesting infection was endemic in the population over the study period. Our results suggested age, pregnancy and lactation were significant risk factors for a detectable neutralizing antibody response. Antibody titres were significantly higher in females than males, with the highest titres occurring in pregnant animals. Temporal variation in antibody titres suggests that herd immunity to the virus may wax and wane on a seasonal basis. These findings support an endemic infection pattern of henipaviruses in bat populations suggesting their infection dynamics may differ significantly from the acute, self limiting episodic pattern observed with related viruses (e.g. measles virus, phocine distemper virus, rinderpest virus) hence requiring a much smaller critical host population size to sustain the virus. These findings help inform predictive modelling of henipavirus infection in bat populations, and indicate that the life cycle of the reservoir species should be taken into account when developing risk management strategies for henipaviruses.     

Measles-virus-specific IgG in optic neuritis and in multiple sclerosis after optic neuritis.

Measles-virus-specific IgG was measured in the serum of 100 patients who had presented with optic neuritis (ON) during 1960-74. When reviewed 41 of them were found to have developed definite symptoms and signs of multiple sclerosis (MS), their serum containing significantly higher titres of the antibody than sera from either the rest of the patients or a group of normal healthy controls. In a few patients from whom cerebrospinal fluid (CSF) was obtained in the acute phase of ON, titres of measles IgG in the serum was higher in those in whom the antibody was detected in the CSF than the serum of patients without CSF antibody.     

Maternally-derived antibodies in pups and protection from canine parvovirus infection

The interaction between maternally-derived antibodies (MDA) and canine parvovirus (CPV) infection was evaluated in five groups of pups with a wide range of haemagglutination inhibiting (HI) titres of MDA (from < 10 to 320). The pups were inoculated with a field CPV strain and monitored daily to evaluate their clinical condition and viral shedding in the faeces. Serum samples were collected weekly to evaluate antibody response. Clinical signs were observed in dogs with HI titres up to 80. Active CPV replication was demonstrated in dogs with HI titres up to 160, although slightly delayed, at lower titres and for a shorter period compared to seronegative dogs. The successful infection of dogs with HI titres of 80 and 160 was also confirmed by seroconversion, evaluated at day 14 postinfection. These findings are in contrast with the MDA titre (HI > or = 80) usually considered fully protective for CPV infection, and suggest the need for revision of current vaccination programmes for pups.     

Infectious pancreatic necrosis virus infection in Atlantic salmon Salmo salar post-smolts affects the outcome of secondary infections with infectious salmon anaemia virus or Vibrio salmonicida.

Infectious pancreatic necrosis (IPN) virus (IPNV) infection in Atlantic salmon Salmo salar L. post-smolts and its influence on the outcome of secondary infections with infectious salmon anaemia (ISA) virus (ISAV) or Vibrio salmonicida were studied. The infections with ISAV or V salmonicida were performed both in a period of acute IPN and in the following IPNV carrier stage, 3 and 6 to 8 wk after experimental IPNV challenge, respectively. An IPNV carrier condition at low virus titre did not influence the mortality rates after secondary infections. Neither the ISAV infection nor the V. salmonicida infection in experimentally induced IPNV carriers resulted in mortalities different from those observed after challenge of IPNV-free fish. At higher IPNV titres in Atlantic salmon with acute IPN, the outcome of secondary infections was quite different from that observed in IPNV-free fish and in IPNV carriers. In 2 different experiments significantly more fish died when fish with acute IPN were infected with V salmonicida than when fish were infected with V salmonicida alone. Mortality also started earlier in the double-infected group than in the group challenged with V. salmonicida alone, 3 to 4 and 8 d after V salmonicida infection, respectively. Similar results were observed independent of whether mortalities due to IPN alone were registered in the experiments. When Atlantic salmon with acute IPN were infected with ISAV, significantly fewer fish died than when fish were infected with ISAV alone. The ongoing IPNV infection seemed to provide some protection against development of ISA.     

Reaginic and homocytotropic IgG antibodies in Schistosoma mansoni infected Mastomys natalensis: time courses in untreated infections and after chemoprophylactic and chemotherapeutic treatment

Mastomys natalensis were infected percutaneously with 250 cercariae of Schistosoma mansoni and treated effectively with 5 X 50 mg amoscanate/kg body-weight 12-16 (I), 28-32 (II), 56-60 (III) and 101-105 days p.i. (IV) respectively. Levels of reaginic antibodies (RAb) and homocytotropic IgG antibodies (GAb) were assessed by passive cutaneous anaphylaxis tests (PCA) and compared with those in infected, untreated animals. In untreated animals RAb were first detected in the 6th week p.i. Maximum titres around 1:80 were reached after 100 days. High levels persisted until the end of the experiment 185 days p.i. RAb did not develop after treatment I. After treatment II only low PCA titres occurred. After treatment III the time course was similar to the controls but levels were reduced. Treatment IV did not interfere with the occurrence of specific RAb. In the case of GAb, independent of any treatment, after a first peak of PCA titres in the 5th week either a negative pattern or only very low titres were found in week 7. In untreated infected Mastomys GAb levels increased thereafter to maximum titres of about 1:80 at day 70. Later than 100 days p.i. relatively constant titres were observed. The second rise of antibody levels did not occur after early treatment. After treatment II only low transient titres developed. Animals treated in the early patency (III) showed reduced PCA titres and were negative at the end of the observation period. Treatment given 101 to 105 days p.i. rather enhanced the development of GAb.     

Enhanced antiplasmin activity in acute renal failure.

Plasmatic slow plasmin-inhibitor activity was assessed in 20 patients with acute renal failure and 12 controls with the fibrin plate method. The area of fibrinolysis was 250-5 +/- 5 mm2 in the patients and 289 +/- 6mm2 in the controls (P less than 0.001) and was negatively correlated with antiplasmin activity. Thirteen patients had areas of lysis equal to or inferior to the minimal lysis observed in the controls. No correlation was found between antiplasmin activity and serum fibrin-fibrinogen related antigen titres, the presence or absence of disseminated intravascular coagulation, or the causative disease.     

Pathology of experimental toxoplasmosis in eastern barred bandicoots in Tasmania

Wild-caught eastern barred bandicoots (Perameles gunnii) initially seronegative to Toxoplasma gondii, were inoculated orally with approximately 100 T. gondii oocysts. The bandicoots were maintained in indoor pens under laboratory conditions and observed daily. Serial blood samples were tested for agglutinating antibodies to T. gondii. Inoculated bandicoots died 15 and 17 days post infection. A rise in Direct Agglutination Test (DAT) titres was detected at the time of death (1:256, 1:64 respectively). Clinical observations, serological changes, gross findings at necropsy, and histopathological changes were consistent with acute toxoplasmosis. The findings indicate that eastern barred bandicoots are likely to die from primary T. gondii infection, often even before detectable antibodies are produced, reinforcing the significance of toxoplasmosis as a potential contributor to the reduction in numbers of wild populations of eastern barred bandicoots.     

Screening of serum antibodies against Mycobacterium tuberculosis by enzyme-linked immunosorbent assay (ELISA) in healthy population

Serum samples of venous blood of healthy individuals, chosen at random from two areas of the People's Democratic republic of Yemen, were examined for the presence of IgG antibodies against mycobacterial antigen by ELISA. From the district of Aden, the capital (A), there were 214 samples (72% vaccinated by BCG, 108 men and 106 women), other 235 ones originated from mountain area Laudar Muhairas (H) (66% vaccinated, 115 men and 120 women). The overall average of acquired titres was 1:81.4 in area A, and an average of 1:102.2 was recorded in area H. There was no substantial difference in the titres level in men of the two areas (1:86.9 and 1:90 resp.), however, a significant difference was observed in women: 1:75.7 in area A and 1:114.7 in area H. In area A the titres were ranging in all age groups from 1:61 to 1:83 and the differences between age groups and between vaccinated and unvaccinated individuals were not significant. In area H the titres were generally higher (1:72 to 1:164) in the unvaccinated and 1:79 to 1:204 in the vaccinated. The differences in the unvaccinated were not of a statistical importance: in the vaccinated, the titres of the 20-29 years old (1:204.5) substantially differed from the titres obtained in other groups. The differences in titres level between the two studied areas are explained by a different epidemiological situation, namely by a higher tuberculosis infestation of some age groups in area H.     

Alternative antigens reduce cross-reactions in an ELISA for the detection of Salmonella enteritidis in poultry

Two alternative antigens for the use in detection of antibodies to salmonellas were investigated: firstly, lipopolysaccharide (LPS) from members of the D2 group, having antigens O : 9, 46, and flagella antigens. Whereas LPS from the D2 group did not discriminate sufficiently with control sera, flagella antigens reacted specifically with antibodies directed to serotype specific H antigens. When flagella antigens were used to screen sera from birds of commercial flocks, however, cross-reactivity between flagella antigens was observed. When both LPS and flagella antigens were used to screen sera from chickens infected with Salmonella, enteritidis, the sera gave higher titres with flagella antigens during the early stages of infection, and titres with flagella antigens dropped earlier after infection had ended than titres with lipopolysaccharide.     

Serologic survey for canine distemper and infectious canine hepatitis in wolves in Alaska

Sera from 57 wolves (Canis lupus) in three areas of Alaska were evaluated for evidence of previous exposure to infectious canine hepatitis virus (ICHV) and canine distemper virus (CDV). Fifty-four sera (94.7%) were positive for ICHV exposure and four (7%) were positive for CDV exposure. All four CDV-reacting wolves also had titres to ICHV. The relatively common occurrence of ICHV exposure may be due to the greater resistance of ICHV to chemical and physical agents and its transmissibility via the urine of infected animals. The ICHV titres observed could indicate enzootic pathogenic ICHV, or exposure to the mildly pathogenic vaccine strain of CAV-1 through contact with the urine of domestic dogs. If CAV-1 is the original source of exposure, the titres could represent an ICHV-protected wolf population.     

Synergy of type I interferon-A6 and interferon-B naked DNA immunotherapy for cytomegalovirus infection

Delivery of type I IFN transgenes by naked DNA immunization can protect against cytomegalovirus infection and myocarditis. Here, we investigate IFN transgene expression, antiviral efficacy, and immunomodulation of myocarditis using various treatment regimes in a mouse CMV model. In vivo expression of the IFN transgene was observed in the sera for 35 days post-DNA inoculation. Prophylactic IFN-A6 and IFN-B DNA treatment for 14 days prior to murine cytomegalovirus (MCMV) infection was more efficacious in significantly reducing viral titres, than 2 days prior to or 2 days post-virus infection. Similarly, IFN-A6 DNA treatment commencing 14 days prior to virus infection was superior in suppressing both acute and chronic myocarditis. Furthermore, reduction of autoantibody titres was more pronounced when IFN was administered 14 days prior to viral infection. Combinational IFN gene therapy was assessed for synergy between IFN subtypes. Combination treatment with either IFN-A6/A9 or IFN-A6/B greatly reduced spleen viral titres while IFN-A6/B and IFN-A9/B reduced virus replication in the liver. Only IFN-A6/A9 and IFN-A9/B reduced acute viral myocarditis, whereas IFNA6/B treatment was most efficacious for autoimmune chronic myocarditis. Finally, treatment with IFN-A6 DNA 2 weeks post-MCMV infection proved effective at inhibiting the development of chronic autoimmune myocarditis. These findings suggest that immunomodulation of both antiviral and autoimmune responses by IFN DNA immunization may be an avenue for improved viral immunotherapy.     

Immunity to equine herpesvirus 1 infection in foals during the first year of life.

A band of 23 pregnant mares on a Thoroughbred breeding farm all had serum virus-neutralizing antibody titres to equine herpesvirus 1 (EHV-1). Antibody was not transferred to their foals in utero. All foals received antibody from colostrum and developed antibody titres similar to their dams. The serum virus-neutralizing antibody titres were observed in 10 of these foals for 1 year. Decay of passive immunity occurred at the rate of 3.25 two-fold dilutions in 100 days and reached zero at the mean time of 180 days. The foals were exposed to EHV-1 twice. Foals with a geometric mean titre of 1 : 25 experienced infection and a rise of titre, while those with a geometric mean titre of 1 : 76 resisted infection.     

A sero-epidemiological study of Leptospira interrogans serovar Balcanica in four brush-tailed possum populations in Victoria, Australia.

The microscopic agglutination (MA) test was utilised to study the prevalence of antibodies to Leptospira interrogans serovar hardjo in 4 populations of brush-tailed possums (Trichosurus vulpecula). The overall antibody prevalence varied from 14% to 66%; however, the age distribution of MA test titres was remarkably similar in all 4 populations. Antibody prevalence was similar in both males and females and demonstrable antibodies were limited to sexually mature animals. The greater prevalence of high titres (greater than or equal to 1:128) in the 18- to 24-month age group suggested that primary infections were acquired at this age. The findings suggested that infection was maintained in possum populations by direct transmission, probably associated with breeding. Focal interstitial nephritis was observed in kidneys of possums greater than 18 months of age and was associated with MA titres to hardjo (P less than 0.001). Serovar balcanica was isolated from possum kidneys from 2 of these populations, suggesting that balcanica infections were responsible for most of the hardjo titres. However, agglutinin-absorption tests indicated that some possums may be infected with a leptospire more closely related to hardjo than to balcanica.     

Kinetics of humoral immune response in pigs vaccinated against foot and mouth disease

The present investigation was conducted to study the foot and mouth disease virus (FMDV)-specific humoral immune response (HIR) in pigs, following vaccination with oil adjuvanted foot and mouth disease (FMD) vaccine, upto 90 days post vaccination (dpv). For this, 40 Large White Yorkshire (LWY) pigs (20; one-year old female (gilts) and 20; three-month old piglets) were vaccinated @ 2 ml/animal, subcutaneously. Sera samples were collected at fortnight interval from all the animals. The log10 SN50 antibody titres against all the serotypes (Type O, A and Asia-1) were detected in both gilts and piglets from day 7 to 90 dpv indicating the persistence of HIR up to the last day of sampling. The maximum antibody titres were observed on 28 dpv, thereafter, titres started declining, but were present till 90 dpv against all the three FMDV serotypes. HIR was more pronounced in piglets in comparison to gilts, as group mean SN antibody titres against all the three FMDV serotypes were found to be more maintained and significantly higher in piglets.     

Thyroxine,methimazole, and thyroid microsomal autoantibody titres in hypothyroid Hashimoto's thyroiditis.

Ten hypothyroid patients with Hashimoto''s thyroiditis were treated with methimazole 30 mg in addition to thyroxine 0.15 mg daily. Another 10 hypothyroid patients with Hashimoto''s thyroiditis were given thyroxine 0.15 mg alone. After 22 weeks of treatment significant decreases in thyroid microsomal autoantibody titres were observed in both groups (p less than 0.01). There was no difference in the mean change in titre between the two groups. When the patients treated with methimazole were subsequently given thyroxine 0.15 mg alone for a further 22 weeks no additional change in titre was observed. The data suggest that thyroxine, by normalising serum thyroid stimulating hormone concentrations, may reduce the autoantigenic properties of the thyrocytes with a subsequent decrease in autoantibody titres.     

The role of IL-12 in the control of MCMV is fundamentally different in mice with a retroviral immunodeficiency syndrome (MAIDS)

The present study investigates the susceptibility of C57BL mice exhibiting T cell immunodeficiency and lymphadenopathy induced by LP-BM5 murine leukaemia virus (MAIDS) to murine cytomegalovirus (MCMV). Treatment of normal (M-) mice with anti-IL-12 increased the contribution of IgG1 to the hypergammaglobulinaemia induced by MCMV, consistent with a shift towards a Th2 phenotype. This impaired control of early MCMV replication in the liver, with little effect in the spleen. Control of hepatic infection correlated with a vigorous splenic NK cytotoxic response in a subgroup of IL-12-depleted M- mice that remained healthy, while others became moribund. Mortality in IL-12-depleted MAIDS (M+) mice given MCMV was ultimately greater than in M- controls, but was delayed despite high levels of MCMV in the liver. IL-12 was required for optimal control of MCMV replication in M+ mice. This may involve cytotoxic activity because similar levels of infection were seen in bg/bg M+ mice, where the beige mutation impairs the formation of cytotoxic granules. Hence the ability of M+ mice to tolerate high titres of MCMV during acute infection may enable innate cytotoxic responses to clear MCMV. Interleukin-12 depletion of M- mice also increased salivary gland MCMV titres and depressed delayed-type hypersensitivity responses to MCMV antigen, normally mediated by CD4+ T cells. These changes were not observed in IL-12-depleted M+ mice.     

Effects of leptin Arg25Cys on peripheral mononuclear cell counts and antibody response to vaccination in beef cattle

A single nucleotide polymorphism (SNP) in the leptin gene that results in Arg25Cys has been associated with beef carcass quality and milk composition in dairy cattle. However, leptin (LEP) also plays a role in immune performance and hence it was important to determine whether selection based on this SNP would negatively affect immune cell numbers or antibody production. LEP c.73C>T was assessed for effects on immune cell counts and antibody titres in 27 beef cattle herds ( n  = 556). A commercial rabies vaccine had been administered to these animals. Prior to being vaccinated, counts of several important mononuclear cells (total and activated B lymphocytes, total and activated T helper and T cytotoxic, WC1 T lymphocytes and monocytes) as well as baseline serum antibody titres were determined for each animal. On day 21, antibody titres were measured and a booster vaccine was administered. Finally on day 42, antibody titres and mononuclear cell types were again counted. Counts of six different cell types were significantly associated with the LEP genotype; however, no consistent patterns were observed between LEP genotype (TT, CT or CC) and peripheral blood mononuclear cell populations. Significant differences in the production of rabies antibodies in response to vaccination were observed relative to LEP genotype. Our results suggest that selection for either the C or T allele would not detrimentally impact on the measured indicators of immune function in beef cattle.     

Comparative immunogenecity of foot and mouth disease virus antigens in FMD-haemorrhagic septicaemia combined vaccine and FMD vaccine alone in buffalo calves

Humoral immune response was evaluated by monitoring the serum antibody titres and virus specific IgM titres against Foot and Mouth Disease (FMD) virus antigens in serum samples obtained from different groups of calves inoculated with combined vaccine or FMD vaccine alone, on 0, 7, 14, 21, 28, 42 and 56 days post-vaccination (DPV). The cellular immune response was monitored by MTT based lymphoproliferation in peripheral blood mononuclear cell cultures. Higher liquid phase blocking (LPB) ELISA antibody titres were observed in calves receiving combined vaccine as compared to calves immunized with FMD vaccine alone with the peak titres in both the groups obtained on 21 days post-vaccination. However, the virus specific IgM titres were significantly higher in group of calves inoculated with combined vaccine than FMD vaccine alone. The lymphoproliferative responses against FMDV types O, A22 and Asia 1 in the groups receiving combined vaccine and FMD vaccine alone started increasing gradually after day 14 and reached peak levels on 28 DPV followed by a gradual decline subsequently. The group receiving combined vaccine showed higher proliferative responses on in vitro stimulation with FMD virus type O, whereas, with FMD virus type Asia 1, the responses were significantly higher on 14 and 21 DPV as compared to the group immunized with FMD vaccine alone. However, in the group receiving combined vaccine, the responses on in vitro stimulation with FMD virus type A22 were significantly higher than FMD vaccine alone group on all DPV except on 42 DPV.     

Postdiarrheal arthropathy of Yersinia pseudotuberculosis.

Two patients with acute gastroenteritis in whom polyarthritis subsequently developed were found to have positive serologic results for Yersinia pseudotuberculosis. With resolution of the arthropathy the antibody titres decreased. While the patient without the histocompatibility antigen HLA-B27 had an acute, self-limited arthritis, the patient with this antigen had a more chronic arthritis. Serologic typing and stool culture for Y. pseudotuberculosis should be done in cases of postdysenteric arthritis.     

Ovarian function in Holstein cows immunized against pregnant mare serum gonadotrophin

Six Holstein-Friesian cows were immunized against pregnant mare serum gonadotrophin (PMSG) using Freunds' adjuvant during the mid-luteal phase of the estrous cycle. Antibody response was maintained by five booster immunizations at 2- to 3-wk intervals. Four cows were treated with a single intramuscular injection of PMSG (2350 I U) 107 d after primary immunization. Cloprostenol (500 ug) was administered at 56 h and 72 h after the treatment with PMSG; the cows were inseminated three times at 12-h intervals starting 56 h after cloprostenol treatment. Five days after insemination, the animals were slaughtered and their reproductive organs were recovered to quantify the population of corpora lutea and unovulated follicles (>10 mm dia). Antibody titres and progesterone concentrations were determined from blood samples collected either on alternate days or twice a week. Initially, progesterone concentrations were measured in milk samples. All cows produced antibodies, and titres were elevated within 6 to 9 d following each booster immunization. After each boost, however, the antibody titres declined rapidly. Progesterone concentrations declined to below 1 ng/ml after two weeks of initial immunization and remained low throughout the study, except in one cow that ovulated on Day 75. All animals were observed to have large follicular cysts during this period. Treatment with PMSG induced a single ovulation in one cow. Ovulations were neither induced by PMSG nor observed in any of the other animals. In PMSG-treated animals, the mean number of large follicles (5.0) was greater than in those which were not treated (2.0). The results of this study suggest that low titres of antibodies against PMSG are sufficient to disturb ovarian activity, result in follicular cysts and block multiple ovulations in response to exogenous PMSG.