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Biomedical subjects

David S Lindsay

Publications and source records attributed to David S Lindsay.

At least 19 recordsLinked to original sources

Effects of high hydrostatic pressure on embryonation of Ascaris suum eggs.

High hydrostatic pressure processing (HPP) has been shown to be an effective non-thermal means of inactivating microorganisms from various food products. Little information is available regarding the effects of HPP on metazoan parasites. Outbreaks of food-borne disease have been associated with importation of food contaminated with fecal material. Ascaris suum is used as a surrogate model metazoan parasite for the human roundworm, Ascaris lumbricoides, to study the effects of treatments on the inactivation of eggs in sludge. The present study was conducted to determine the effects of HPP on A. suum eggs. Unembryonated A. suum eggs were subjected to 138-552 megapascals (MPa) for 10-60s in a commercial HPP unit. Embryonation was induced after HPP treatments by incubating eggs in 0.01N sulfuric acid at room temperature. After 21 days, 100 eggs were examined per treatment using a light microscope and the percent of embryonated eggs was determined. Embryonation was induced in 38-76% eggs that were subjected to 138 and 270MPa. No embryonation was observed in eggs exposed to pressures of 241MPa or more for 60s or in eggs exposed to 276MPa for 10-30s. These results indicate that HPP treatment could be used to protect contaminated food items by inactivating A. suum eggs and may also have potential in reducing food-borne illness resulting from fecal contamination.

Animals↗

Seroprevalence of Toxoplasma gondii, Sarcocystis neurona, and Encephalitozoon cuniculi in three species of lemurs from St. Catherines Island, GA, USA.

In the current study, we determined the seroprevalence of Toxoplasma gondii, Sarcocystis neurona, and Encephalitozoon cuniculi in three species of lemurs from St. Catherines Island, Georgia. Serum samples were tested from 52 ring-tailed lemurs (Lemur catta), six blue-eyed black lemurs (Eulemur macaco flavifrons), and four black and white ruffed lemurs (Varecia variegata variegata) using an agglutination assay. Three ring-tailed lemurs (5.8%) were positive for T. gondii (titer of 1:50); one ring-tailed lemur (1.9%) and one black and white ruffed lemur (25%) were positive for S. neurona (titers of 1:1000); and one ring-tailed lemur (1.9%) was positive for E. cuniculi (titer of 1:400). All blue-eyed black lemurs were negative for antibodies to T. gondii, S. neurona, and E. cuniculi. This is the first detection of antibodies to T. gondii in ring-tailed lemurs and antibodies to S. neurona and E. cuniculi in any species of prosimian.

Agglutination Tests↗

Prevalence of antibodies to Encephalitozoon cuniculi in horses from Brazil.

Encephalitozoon cuniculi has been associated with natural cases of abortion and stillbirth in horses. However, little is known about the prevalence of this parasite in horses. We examined sera from 559 horses from Brazil for antibodies to E. cuniculi using the indirect immunofluorescent antibody (IFA) test and the direct agglutination test (DAT). We found that 79 (14.1%) were positive in the IFA test and 70 (12.5%) were positive in the DAT. Compared to the IFA as the "gold standard" the sensitivity of the DAG was 94.0% and the specificity was 96.1%. Our study indicates that horses in Brazil are frequently exposed to E. cuniculi.

Agglutination Tests↗

Immune response to Sarcocystis neurona infection in naturally infected horses with equine protozoal myeloencephalitis.

Equine protozoal myeloencephalitis (EPM) is one of the most common neurologic diseases of horses in the United States. The primary etiologic agent is Sarcocystis neurona. Currently, there is limited knowledge regarding the protective or pathophysiologic immune response to S. neurona infection or the subsequent development of EPM. The objectives of this study were to determine whether S. neurona infected horses with clinical signs of EPM had altered or suppressed immune responses compared to neurologically normal horses and if blood sample storage would influence these findings. Twenty clinically normal horses and 22 horses with EPM, diagnosed by the presence of S. neurona specific antibodies in the serum and/or cerebrospinal (CSF) and clinical signs, were evaluated for differences in the immune cell subsets and function. Our results demonstrated that naturally infected horses had significantly (P<0.05) higher percentages of CD4 T-lymphocytes and neutrophils (PMN) in separated peripheral blood leukocytes than clinically normal horses. Leukocytes from naturally infected EPM horses had significantly lower proliferation responses, as measured by thymidine incorporation, to a non-antigen specific mitogen than did clinically normal horses (P<0.05). Currently, studies are in progress to determine the role of CD4 T cells in disease and protection against S. neurona in horses, as well as to determine the mechanism associated with suppressed in vitro proliferation responses. Finally, overnight storage of blood samples appears to alter T lymphocyte phenotypes and viability among leukocytes.

Animals↗

Penetration of equine leukocytes by merozoites of Sarcocystis neurona.

Horses are considered accidental hosts for Sarcocystis neurona and they often develop severe neurological disease when infected with this parasite. Schizont stages develop in the central nervous system (CNS) and cause the neurological lesions associated with equine protozoal myeloencephalitis. The present study was done to examine the ability of S. neurona merozoites to penetrate and develop in equine peripheral blood leukocytes. These infected host cells might serve as a possible transport mechanism into the CNS. S. neurona merozoites penetrated equine leukocytes within 5 min of co-culture. Infected leukocytes were usually monocytes. Infected leukocytes were present up to the final day of examination at 3 days. Up to three merozoites were present in an infected monocyte. No development to schizont stages was observed. All stages observed were in the host cell cytoplasm. We postulate that S. neurona merozoites may cross the blood brain barrier hidden inside leukocytes. Once inside the CNS these merozoites can egress and invade additional cells and cause encephalitis.

Animals↗

Neosporosis, toxoplasmosis, and sarcocystosis in ruminants.

Neospora caninum, Sarcocystis spp, and Toxoplasma gondii are related coccidian parasites that can cause abortion and neonatal mortality in animals. In addition, T gondii and certain species of Sarcocystis are zoonotic. This article reviews information on the etiology, diagnosis, control, and prevention of these diseases.

Animals↗

Identification of new morphological and life-cycle stages of Cochlosoma anatis and experimental transmission using pseudocyst.

Cochlosoma anatis is a flagellated intestinal parasite that infects a variety of avian species. C. anatis infections have been associated with decreased weight gain and increased morbidity and mortality. Conditions favoring the growth of this organism in birds are current pathogenic intestinal infections and/or young age. There is little data describing the life cycle of this parasite. In this study, electron microscopy images are presented that document longitudinal binary fission of the trophozoite stage and outline the events of pseudocyst formation, which includes a rounding stage. Evidence provided here indicates that the pseudocyst stage may be a mechanism for transmission of this organism. The observations reported here provide additional evidence of homology between Cochlosoma and members of the trichomonad order.

Animals↗

Effects of high-pressure processing on Toxoplasma gondii tissue cysts in ground pork.

Ingestion of Toxoplasma gondii tissue cysts can result in severe disease in immunocompromised individuals and pregnant women. Treatment of meat and meat products to eliminate viable T. gondii tissue cysts would provide a means to protect consumers. In this study, we examined the effects of high-pressure processing (HPP) on ground pork containing viable tissue cysts of the VEG strain of T. gondii. Ground pork containing tissue cysts was exposed to 400, 300, 200, 100, or 0 MPa treatment for 30, 60, or 90 sec in a commercial HPP unit. The HPP-treated ground pork was subjected to acid-pepsin digestion and bioassayed in mice. The results of the mouse bioassay revealed that none of the mice inoculated with tissue cysts exposed to 400 or 300 MPa became infected, whereas all mice inoculated with tissue cysts exposed to 200, 100, or 0 MPa became infected with T. gondii regardless of exposure time. Results indicate that HPP treatment of ground pork with 300 MPa of pressure will render tissue cysts of T. gondii nonviable and make pork safe for human consumption.

Air Pressure↗

Prevalence of agglutinating antibodies to Toxoplasma gondii in striped skunks (Mephitis mephitis), opossums (Didelphis virginiana), and raccoons (Procyon lotor) from Connecticut.

The prevalence of agglutinating antibodies to Toxoplasma gondii was examined in striped skunks (Mephitis mephitis), opossums (Didelphis virginiana), and raccoons (Procyon lotor) from 8 cities in Connecticut. Ten (42%) of the 24 striped skunks, 2 of 7 (29%) opossums, and 12 of 12 (100%) raccoons were positive at dilutions of 1:50 or greater. These results suggest that T. gondii is prevalent in the environment, or prey items, or both, of these omnivores in Connecticut.

Agglutination Tests↗

Activity of bleach, ethanol and two commercial disinfectants against spores of Encephalitozoon cuniculi.

Encephalitozoon cuniculi is a small protist parasite in the phylum Microspora. Hosts are infected by ingestion or inhalation of spores passed in the urine or feces. Infection with E. cuniculi is usually asymptomatic, except in young or immunocompromised hosts. This study examined the effects of various disinfectants on in vitro infectivity of E. cuniculi spores. Spores of E. cuniculi were exposed to several dilutions of commercial bleach, 70% ethanol and dilutions of commercial disinfectants HiTor and Roccal for 10 min and then loaded onto human fibroblast cells (Hs68 cells). Ten minutes of exposure to these disinfectants was lethal to E. cuniculi spores. Additional exposure time studies were done using dilutions of bleach at 0.1, 1 and 10%, and 70% ethanol. Exposure of E. cuniculi spores to 1 or 10% bleach for 30s rendered them non-infectious for Hs68 cells. Growth of E. cuniculi was observed in Hs68 cells inoculated with spores treated with 0.1% bleach for 30s or 1, 3 and 5 min, but not with spores treated for 7 min or longer. Exposure of E. cuniculi spores to 70% ethanol for 30s rendered them non-infectious for Hs68 cells. Spores of E. cuniculi are more sensitive to disinfectants than are coccidial oocysts and other parasite cysts. The relatively short contact time needed to kill spores indicates that disinfection of animal housing may be a viable means to reduce exposure of animals to E. cuniculi spores.

Animals↗

Direct agglutination test for Encephalitozoon cuniculi.

Encephalitozoon cuniculi is a small protozoan parasite in the phylum Microspora. It has been shown to naturally infect several host species, including humans. Infection with microsporidia is usually asymptomatic, except in young or immunocompromised hosts. Currently, serological diagnosis of infection is made using the indirect immunofluorescent antibody assay (IFA) or enzyme-linked immunosorbent assay (ELISA). Although these methods are sensitive and reliable, there are several drawbacks to the IFA and ELISA tests. Cross-reactivity between other Encephalitozoon species is common, and specialized equipment is required to conduct these tests. This paper reports the development of a direct agglutination test for detecting IgG antibodies to E. cuniculi. The utility of the agglutination test was examined in CD-1 and C3H/He mice infected with E. cuniculi or one of 2 other Encephalitozoon species. Test sera were incubated overnight with eosin-stained microsporidia spores in round-bottom microtiter plates. In positive samples, agglutination of spores with antibodies in test sera resulted in an opaque mat spread across the well. The results indicate that the agglutination test is 86% sensitive and 98% specific for E. cuniculi, with limited cross-reactivity to Encephalitozoon intestinalis. No cross-reactivity to Encephalitozoon hellem was observed. The test is fast and easy to conduct, and species-specific antibodies are not required.

Agglutination Tests↗

Placentitis associated with leishmaniasis in a dog.

A 1.5-year-old Coonhound from Maryland aborted 7 fetuses. Placenta and internal tissues of 1 fetus were examined histologically. The predominant lesion was placentitis characterized by necrosis and infiltration of mixed leukocytes. Numerous Leishmania spp amastigotes were identified in placental trophoblasts, and the diagnosis was confirmed by use of immunohistochemical staining with Leishmania-specific antibodies. Protozoa were not found in the fetal tissues. An indirect fluorescent antibody test yielded a serum titer of 1:100, and a recombinant K39 immunoassay of serum yielded positive results for the K39 Leishmania antigen.

Abortion, Veterinary↗

Flow cytometric analysis of cellular immune responses in dogs experimentally infected with a North American isolate of Leishmania infantum.

Canine leishmaniasis caused by Leishmania infantum is endemic in the foxhound population in North America. Studies of canine leishmaniasis in the Mediterranean basin indicate a role for both CD4+ and CD8+ lymphocytes with clinical illness and in asymptomatic dogs. Limited information is available on the strain of L. infantum infecting foxhounds in North America. The present study investigated changes in cellular immune responses in dogs experimentally infected with 1x10(7) (low dose, LD; N=4) or 2x10(8) (high dose, HD; N=4) promastigotes of a United States isolate of L. infantum and control dogs (N=2) for 72 weeks. Density gradient separation was used to enrich for peripheral blood lymphocytes from canine blood. Lymphocyte subsets (CD4+ and CD8+) were quantified by flow cytometric analysis. Lymphocyte population expression levels over the course of the present study were compared to clinical status of the dog and antibody responses in infected and control dogs. No significant differences (P>0.05) were observed in either CD4+ or CD8+ lymphocyte expression in of the groups over the experimental period. This study suggests that the cellular immune responses to North American L. infantum in experimentally infected dogs may differ from other strains of L. infantum.

Animals↗

Infections in immunocompetent and immune-deficient mice with promastigotes of a North American isolate of Leishmania infantum.

Leishmania infantum, an etiologic agent of zoonotic visceral leishmaniasis, is widespread among foxhounds in the United States. Experimental infections with a North American isolate of L. infantum were evaluated using two inoculation routes in immunocompetent and immunosuppressed mouse strains. Groups of 2-5 interferon gamma gene knockout (IFN-gamma-KO) (BALB/c-Ifng), inducible nitric oxide synthase (NOS) gene knockout (iNOS-KO) (C57BL/6), B-cell-deficient (microMT) (C57BL/6), and BALB/c mice were intravenously (i.v.) or subcutaneously (s.c.) inoculated with various doses of promastigotes of the LIVT-1 strain of L. infantum. None of the mice developed clinical signs of leishmaniasis during the 8-9 weeks of the study. Promastigotes were cultured from spleens of all i.v.-infected mice by 3 days post culture. Spleens from s.c.-infected mice inoculated with greater than 1 x 10(6) parasites became culture positive 3-24 days post culture, but promastigotes were not cultured from mice infected with 1 x 10(5) or 5 x 10(5) LIVT-1 promastigotes. Histological lesions were prominent in the livers of i.v.-infected mice but were mild to nonexistent in s.c. infection. Serological responses were low and transient determined by indirect fluorescent antibody testing in all groups. These results indicate that the i.v. route of infection is superior to the s.c. route in a mouse model of North American leishmaniasis and that mice lacking INF-gamma, iNOS or mice that are B-cell-deficient are not more susceptible to acute infection.

Animals↗

Gerbil model of acute neosporosis.

Experimental infections with the NC-1 strain of Neospora caninum were conducted in gerbils (Meriones unguiculatus) to determine their acute responses to experimental intraperitoneal infection. Five groups of five female gerbils were used and they were intraperitoneally infected with 1x10(6), 2x10(6), 3x10(6), 4x10(6) or 5x10(6) tachyzoites. Gerbils in all groups developed clinical signs of neosporosis which consisted of inactivity 4-5 days post-inoculation. Morbidity and mortality were observed in all groups. Grossly there was a clear fibrinous exudate in the abdominal cavity and adhesions of the spleen and pancreas to the stomach in gerbils suffering from acute neosporosis. The LD50 was calculated as 9.3x10(5) tachyzoites per gerbil. The results indicate that gerbils can be used as a suitable model of acute neosporosis. This model can be used to screen candidate treatments and to test the efficacy of vaccines for neosporosis without the need to use histology or PCR to demonstrate treatment efficacy.

Acute Disease↗