Enhancement of risk of tick-borne infection: environmental and parasitological aspects of the problem

The appearance of phenotypic (and probably genetic) exoskeleton anomalies in Ixodes ticks, first discovered and described elsewhere, seems to be a global phenomenon clearly related to environment pollution by heavy metal ions. These external markers of cadmium accumulation in the tick indicate an enhanced risk of tick-borne infection related to an increased vectorial capacity. This manifests itself in the ability of infected ticks both to quest longer and to show increased locomotory activity (hunting) compared with normal ticks. Ticks with exoskeleton anomalies show a greater susceptibility to different microorganisms, including tick-borne pathogens, and more intense pathogen replication with a higher prevalence of multi-infection.     

Experimental infection of pigs with West Nile virus

Summary. Young adult and weanling pigs were challenged with the New York 99 strain of West Nile virus through the bite of infected mosquitoes. Each of six adult pigs seroconverted, but virus was isolated from serum of only one pig following challenge. Three of five weanling pigs developed viremia, with peak titers of 10^1.9 and 10^3.1PFU/mL. Clinical signs attributable to West Nile virus infection were not observed in any of these animals. An additional four pigs were challenged by feeding West Nile virus-infected mice, and none of the four developed a detectable viremia or seroconverted. These results suggest that pigs are unlikely to play a significant role as amplifying hosts of West Nile virus.     

Experimental infection with Treponema hyodysenteriae in guinea pigs.

Outbred and inbred (Hartley strain) guinea pigs (GP) were inoculated intragastrically with pathogenic and nonpathogenic Treponema hyodysenteriae. GP 3 to 16 weeks old received T. hyodysenteriae after a fasting period of 36 to 72 h. Infected GP with pathogenic T. hyodysenteriae developed a diarrheal and/or depressive condition, with mucus but not blood in the feces. Of 88 GP, 40 had gross lesions resembling those of swine dysentery. Lesions were limited mainly to the large intestine. TP used as controls or inoculated with nonpathogenic T. hyodysenteriae did not develop these lesions in the large intestine. These studies suggest that the GP may be used as an animal model for swine dysentery.     

Experimental Nipah virus infection in pigs and cats

A human isolate of Nipah virus from an outbreak of febrile encephalitis in Malaysia that coincided with a field outbreak of disease in pigs was used to infect eight 6-week-old pigs orally or subcutaneously and two cats oronasally. In pigs, the virus induced a respiratory and neurological syndrome consistent with that observed in the Malaysian pigs. Not all the pigs showed clinical signs, but Nipah virus was recovered from the nose and oropharynx of both clinically and sub-clinically infected animals. Natural infection of in-contact pigs, which was readily demonstrated, appeared to be acute and self-limiting. Subclinical infections occurred in both inoculated and in-contact pigs. Respiratory and neurological disease was also produced in the cats, with recovery of virus from urine as well as from the oropharynx. The clinical and pathological syndrome induced by Nipah virus in cats was comparable with that associated with Hendra virus infection in this species, except that in fatal infection with Nipah virus there was extensive inflammation of the respiratory epithelium, associated with the presence of viral antigen. Viral shedding via the nasopharynx, as observed in pigs and cats in the present study, was not a regular feature of earlier reports of experimental Hendra virus infection in cats and horses. The findings indicate the possibility of field transmission of Nipah virus between pigs via respiratory and oropharyngeal secretions.     

Streptococcus suis serotype 2 infection in pigs: new diagnostic and pathogenetic aspects

In a study aimed at improving the diagnosis and elucidating the pathogenesis of Streptococcus suis serotype 2 infection in pigs, a combination of bacterial culture and histopathological and immunohistochemical examination was applied to a range of tissues from 42 naturally infected pigs with typical macroscopical lesions. By culture, 21 pigs (50%) were shown to be systemically infected with S. suis serotype 2; seven (17%) were infected with S. suis serotype 7, two with other bacteria, and 12 yielded no bacterial pathogens. The highest isolation rate for S. suis serotype 2 was obtained from the lateral cerebral ventricles and other regions of the brain, whereas the bacterium was only rarely isolated from the liver or spleen. Immunohistochemically, a diagnosis of S. suis serotype 2 infection was obtained in two of 12 (17%) animals from which no pathogens had been cultured. Moreover, immunohistochemistry differed from culture in revealing a greater number of positive tissue specimens. The microanatomical distribution of bacteria pointed toward the pharyngeal and palatine tonsils as principal portals of entry. Furthermore, S. suis serotype 2 bacteria were frequently identified immunohistochemically in the regional lymph nodes of the upper respiratory tract, possibly reflecting primary lymphogenous spread from the tonsils.     

Experimental infection and immune response of guinea pigs with varicella-zoster virus.

An immune response (fluorescent antibody to membrane antigen) was detected in guinea pigs inoculated with varicella-zoster virus (VZV) adapted to guinea pig embryonic cells, including the Oka vaccine strain, even when inoculation was by an external route, i.e., nasal or corneal. Live or UV-inactivated virus having the same virus titer before irradiation was administered to guinea pigs by the corneal route, and antibody induction was detected only with live virus. The transmission of VZV from infected guinea pigs to noninfected ones was suggested by the appearance of antibody in the serum of the latter, who were kept in the same cage. The time course of the appearance of humoral and cellular immune responses in guinea pigs was examined by the fluorescent antibody to membrane antigen test and the skin reaction, with varicella antigen representing delayed-type hypersensitivity. When VZV was injected subcutaneously, skin reaction appeared as early as 4 days after inoculation, which preceded the appearance of detectable antibody by 2 to 6 days. In in vitro studies, the Oka vaccine showed a higher adsorption rate and better growth in guinea pig embryonic cells than did other wild-type strains when assayed by the infectious center assay. These results suggest that a system of VZV adapted to guinea pig cells and guinea pigs provides a good animal experimental model for immunological study of VZV infection.     

Evaluation of parasitological and immunological aspects of acute infection by Ancylostoma caninum and Ancylostoma braziliense in mixed-breed dogs

This study compared the course of infection by Ancylostoma caninum and Ancylostoma braziliense in mixed-breed dogs infected with L₃ larvae. Dogs infected with A. caninum eliminated more eggs than did those infected with A. braziliense. A total of 38 % of A. caninum and 44 % of A. braziliense larvae were recovered as adult worms. There were no marked clinical abnormalities in dogs with either infection. A. caninum was associated with anemia and an increased number of circulating neutrophils, whereas infection with A. braziliense led to a decrease in the number of leukocytes. The humoral response against excreted and secreted antigens from adult worms was more sensitive and specific than the response induced with the crude antigen. No immune response was observed for either crude or excreted-secreted (ES) antigens from larvae of either species. A nonspecific response against the crude antigen of A. braziliense was found at 0 and 7 days postinfection and maintained throughout the infection period. However, antibody titers against ES antigens were elevated in A. caninum infection at patency and death, showing that this antigen has a higher specificity. The immune response elicited by infection with A. braziliense in dogs has not been described previously. No significant differences were observed in the infection processes of the two Ancylostoma species, except for the higher number of eggs eliminated from dogs infected with A. caninum, which may indicate a better evolutionary adaptation of the parasite to its host in comparison with A. braziliense.     

Experimental infection of laying turkeys with Rhinotracheitis virus: Distribution of virus in the tissues and serological response

Twenty-four laying turkey hens shown to be free of antibodies to turkey rhinotracheitis virus were inoculated intranasally with an isolate of the virus. A mild respiratory disease developed between 5 and 9 days post infection (pi). Two birds were selected at random at intervals between days 1 and 20 pi, killed and tissues examined for the presence of virus. At autopsy between days 2 and 12 abnormalities were found in the oviducts including the deposition of inspissated albumen. Yolk material was occasionally found in the abdominal cavity and there was one instance of egg peritonitis. Eggs with abnormal shells were found in the uterus on days 3 and 9. By direct immunofluorescence (IF) staining, virus was detected in the trachea between days 1 and 7 pi and in the turbinates between days 2 and 5 pi. Virus could also be isolated from these sites using turkey embryo tracheal organ cultures but this method was slightly less sensitive than IF for these tissues. No virus was demonstrated in the lungs or air sacs. Viral antigens were detected by IF in the epithelium of the uterus on day 7 pi and in this and all other regions of the oviduct on day 9 pi. Virus was isolated only from middle magnum and vagina on day 9 pi. On other occasions up to 20 days pi the above tissues and spleen, ovary, liver, kidney and hypothalamus were all negative for virus. Antibodies detected by ELISA and serum neutralisation both reached, high titre by 12 days pi and were maintained at a high level (Iog2 12-15) throughout the period of observation (89 days).     

Insecticide resistance in head lice: clinical,parasitological and genetic aspects

Insecticide treatment resistance is considered to be a major factor in the increasing number of infestations by head lice. The large insecticide selection pressure induced by conventional topical pediculicides has led to the emergence and spread of resistance in many parts of the world. Possible mechanisms of resistance include accelerated detoxification of insecticides by enzyme-mediated reduction, esterification, oxidation that may be overcome by synergistic agents such as piperonyl butoxide, alteration of the binding site, e.g. altered acetylcholinesterase or altered nerve voltage-gated sodium channel, and knockdown resistance (kdr). Clinical, parasitological and molecular data on resistance to conventional topical pediculicides show that treatments with neurotoxic insecticides have suffered considerable loss of activity worldwide. In particular, resistance to synthetic pyrethroids has become prominent, probably because of their extensive use. As other treatment options, including non-insecticidal pediculicides such as dimeticone, are now available, the use of older insecticides, such as lindane and carbaryl, should be minimized, owing to their loss of efficacy and safety concerns. The organophosphorus insecticide malathion remains effective, except in the UK, mostly in formulations that include terpineol.     

Chronic urticaria: novel clinical and serological aspects

BACKGROUND: Recently, distinct studies have shown that: (a) chronic idiopathic urticaria (CIU) is autoimmune in 30-50% of cases; (b) in patients with CIU the autologous serum skin test is inhibited by heparin; and (c) basophil histamine release induced in vitro by CIU sera maybe complement-dependent. OBJECTIVE: To carry out a comprehensive clinical and serological study on CIU based upon these observations. METHODS: Three hundred and six adults with CIU underwent intradermal (ID) test with autologous serum; 57 of them with autologous heparinized plasma as well. Sera from 121 patients (plasmas from 17) were employed to induce in vitro histamine release from basophils of normal donors. The effects of heating (56 degrees C, 60 min), filtration through membrane, and preincubation with heparin were evaluated as well. RESULTS: Autologous serum and plasma induced a weal and flare reaction in 205 out of 306 (205/306; 67%) and in 8/57 (14%) patients, respectively. Positive plasma skin tests were observed only in patients showing strongly positive serum skin tests. Plasma did not elicit any skin reaction in 3/3 patients with dermatographism who showed a positive intradermal test with saline. Sera from 20/121 (16.5%) patients induced significant histamine release from basophils of normal donors. 19/20 sera were from patients with a positive intradermal test; thus, basophil histamine release assay was positive in 19/87 (21.8%) patients with a positive serum skin test. Heating at 56 degrees C x 1 h markedly reduced the histamine-releasing activity of both serum and plasma from in vitro reactors. Ultrafiltered fractions > 100 kDa of both sera tested retained the histamine-releasing activity, whereas fractions < 100 kDa were not able to induce any histamine release. Heparin dose-dependently inhibited histamine release induced by sera and plasma, and by basophil agonists such as anti-IgE, formyl-methionyl-leucyl-phenilalanine, and interleukin (IL)-3. CONCLUSIONS: 67% of our patients with CIU showed a positive autologous serum skin test. Sera from about 20% of those positive on autologous serum skin test induced histamine release from normal basophils in vitro probably as a consequence of the presence of functional autoantibodies. The marked difference between in vivo and in vitro findings could reflect the existence of a mast cell-specific histamine-releasing factor which does not release histamine from basophils of healthy blood donors. However, it might be also the result of in vivo priming of patients' cutaneous mast cells or of heterogeneity of basophil donors. At least in some cases complement seems essential for histamine-releasing activity of serum from patients with CIU. Heparin inhibits histamine release from both basophils (in vitro) and mast cells (in vivo), probably acting directly at a cellular level.     

Experimental in utero infection of fetal pigs with a porcine parvovirus.

In utero infection of fetuses of six specific-pathogen-free large white sows at 35, 48, 55, 72, 99, and 105 days was studied. The fetuses were infected by direct inoculation of porcine parvovirus into the amniotic sac. The inoculation consisted of 0.25 ml of tissue culture fluid containing 10(5.5) mean tissue culture infective doses per ml of porcine parvovirus strain G10/1. Fetuses of one uterus horn were infected, whereas fetuses in the opposite horn were given 0.25 ml of noninfected cell culture material. No clinical signs of infection were observed; however, all sows developed antibodies 7 to 9 days postinfection. A total of 24 virus-inoculated fetuses and 20 control fetuses were studied. Fetuses infected at 35, 48, and 55 days of gestation died between about 5 and 22 days after infection. Virus was isolated from their organs and fetal blood. Virus spread to control fetuses but did not cause death and mummification or stimulate antibody production. Fetuses from sows infected at 72, 99, and 105 days of gestation survived. They developed high antibody titer in utero. Control piglets remained antibody free.     

Experimental Helicobacter pylori gastric infection in miniature pigs

An experimental Helicobacter pylori infection in miniature pigs was developed and investigated. Eighteen miniature pigs were inoculated with an H. pylori strain that has high virulence in mice at c. 5 x 10(10) cfu. H. pylori infection in miniature pigs was achieved by the administration of agar 1% in brucella broth with fetal bovine serum 10% just before inoculation. The bacterial colonisation and distribution were analysed by mapping of viable cell counts in the stomach in pigs of three different ages. The mapping assay was achieved on post-infection day 3 for the 5-day-old and 2-week-old pigs, and between days 41 and 43 for 3-month-old pigs. The highest cell counts were observed in 5-day-old pigs, which averaged 4.9 x 10(6) cfu/g of mucosa (n = 4). The bacteria were colonised mainly in the cardiac and fundus gland region in the 5-day-old and 2-week-old pigs, whereas the colonisation sites did not depend on the region in the 3-month-old pigs. Biopsy assay of the antral mucosa of a 3-month-old pig after H. pylori infection showed that this infection persisted for >22 months. Serum antibody against H. pylori was detected in the infected pigs but not in the uninfected animal. Immunostaining demonstrated the presence of bacteria on the epithelial surface of the infected pigs. A microscopic finding common to all the infected pigs, focal gastritis with infiltration of lymphocytes detected on the lesser curvature of the stomach, resembled the microscopic appearance in H. pylori-infected human patients. These results suggest that miniature pigs might be a suitable model for studying H. pylori infection.     

Experimental infection of newly weaned pigs with human and porcine strains of Serpulina pilosicoli.

Cultures of Serpulina pilosicoli 95/1000, isolated from a pig with porcine intestinal spirochetosis (PIS), and S. pilosicoli WesB, isolated from an Aboriginal child with diarrhea, were used to infect 5-week-old newly weaned pigs. Four of 12 pigs infected with strain 95/1000 and 2 of 12 pigs infected with strain WesB became colonized and developed watery, mucoid diarrhea within 2 to 11 days postinfection. Affected pigs all had moderate subacute mucosal colitis, with gross and histological changes similar to those previously reported in both natural and experimentally induced cases of PIS. Silver-stained histological sections of the colon and cecum from affected pigs demonstrated spirochetes within dilated intestinal crypts, where they were associated with neutrophilic exocytosis and mucus secretion. Sections from one pig infected with strain 95/1000 showed large numbers of spirochetes attached by one end to the colonic epithelium, a feature consistent with PIS. This study confirms the role of S. pilosicoli in the etiology of PIS and provides evidence that S. pilosicoli strains of human origin have pathogenic potential in an animal model.     

Experimental inoculation of conventional pigs with tissue homogenates from pigs with post-weaning multisystemic wasting syndrome.

This report describes the experimental inoculation of conventional pigs with a tissue homogenate obtained from two pigs affected with postweaning multisystemic wasting syndrome (PMWS). Eight 2-month-old pigs were inoculated by the intranasal route, and two pigs were left as uninfected controls. Clinical signs, rectal temperatures and body weights were recorded. Pigs were necropsied at days 14 or 21 post-inoculation, and tissue samples were taken for histopathology and porcine circovirus (PCV) in-situ hybridization. Although only mild clinical signs of disease were observed, lesions of PMWS were seen, and PCV was shown to have been successfully transmitted to six of the eight pigs. Seroconversion of all inoculated pigs to PCV-2, but not to PCV-1, was also detected, suggesting that the PCV nucleic acid detected by in-situ hybridization in inoculated pigs corresponded to PCV-2. In conclusion, this report shows that PCV-2 is transmissible to pigs, and the inoculation of tissue homogenates containing the virus results in the development of PMWS-like lesions.