scholarly journals Neospora caninum DNA distribution in tissues of gerbils as experimental models of chronic neosporosis

2020 ◽  
Vol 72 (5) ◽  
pp. 1719-1726
Author(s):  
G. Toscan ◽  
A.S. Cezar ◽  
P. Bräunig ◽  
G.R. Pereira ◽  
A.C. Vargas ◽  
...  

ABSTRACT Neospora caninum is the main etiologic agent of neosporosis in domestic animals and its pathogenesis comprises two characteristic phases: acute and chronic. Rodents are used as experimental models to mimic acute and chronic bovine neosporosis. In this study, we inoculated a total of 27 female gerbils, with different doses of N. caninum tachyzoites aiming to induce chronic disease. DNA was extracted from different organs of each animal after spontaneous death or euthanasia. Encephalic tissues were submitted to a highly sensitive real time PCR aiming to detect chronically infected animals. All the other samples were submitted to standard PCR. A total of 11 gerbils died due to acute neosporosis, as confirmed by N. caninum DNA detection in organs. 5x103 tachyzoites/mL of N. caninum was the dosage of antigen that can induce chronic infection in gerbils. In the encephalon sections of some animals that showed clinical signs of persistent infection, we found 70% positive for the anterior encephalon section, suggesting this area as preferential for cyst formation. Therefore, we determined the doses of tachyzoites that cause acute or chronic infection and detection of positive tissues, preferably, systemic organs during acute and encephalon in chronic phases.

1990 ◽  
Vol 68 (7) ◽  
pp. 1595-1599 ◽  
Author(s):  
David S. Lindsay ◽  
J. P. Dubey

The susceptibility of laboratory rats (Rattus norvegicus) to experimental inoculation with tachyzoites of Neospora caninum was examined. Groups of female rats were intramuscularly injected with 0, 2, or 4 mg of methylprednisolone acetate (MPA) 7 days prior to, and on the day of, subcutaneous inoculation with 0 or 1.5 × 105 tachyzoites. Clinical signs of disease or deaths did not occur in rats given nothing or only N. caninum tachyzoites. Rats given only 4 mg MPA failed to grow as well as rats given nothing or only N. caninum tachyzoites but were otherwise healthy. All of 20 rats given 4 mg MPA and tachyzoites died of hepatitis and pneumonia within 12 days postinoculation. Hepatic necrosis was the most striking lesion seen in these rats, and other milder lesions consisted of pneumonia, encephalitis, and myositis. The response of rats given 2 mg MPA and tachyzoites was less severe. Three of 20 rats died with encephalitis, myositis, hepatitis, and pancreatitis. Mild lesions, but no N. caninum tachyzoites, were seen in 3 of 14 rats inoculated only with tachyzoites. Rats given the 4 mg MPA treatment and inoculated with N. caninum tachyzoites appear to be suitable subjects for examining acute neosporosis and could be used in studies designed to examine treatment of acute disease.


1996 ◽  
Vol 33 (6) ◽  
pp. 647-655 ◽  
Author(s):  
M. M. McAllister ◽  
A. M. McGuire ◽  
W. R. Jolley ◽  
D. S. Lindsay ◽  
A. J. Trees ◽  
...  

Six groups of six pregnant ewes each were inoculated with 170,000 or 1,700,000 tachyzoites of Neospora caninum on gestation day 65, 90, or 120. All ewes seroconverted, and none showed signs of illness other than abortion. Regardless of the inoculum dose, all ewes inoculated on gestation day 65 aborted: ewes inoculated on gestation day 90 aborted, gave birth to weak lambs, or gave birth to clinically normal lambs; and all ewes inoculated on gestation day 120 gave birth to clinically normal lambs. Using an immunohistological procedure that stains bradyzoites, we observed protozoal cysts in brains of 11 of 29 (38%) aborted fetuses, in one of four (25%) weak lambs, and in seven of 18 (39%) clinically normal lambs. Cysts were not observed in extraneural tissues from two clinically normal lambs that had cysts in the brain. No evidence of infection was observed in tissues of five ewes examined using an immunohistological procedure that stains N. caninum tachyzoites and bradyzoites. Multifocal nonsuppurative encephalitis was observed in 46 of 51 (90%) aborted, weak, or clinically normal lambs. Cerebral necrosis, dystrophic mineralization, and meningitis were also commonly identified in live and aborted lambs (even when severely autolyzed). Nonsuppurative, necrotizing placentitis was observed in 15 of 17 (88%) placentas. Nonsuppurative myositis was common in fetuses but not in live lambs. Inflammation occurred less frequently in liver and lung. Clinical and pathological features of neosporosis in sheep closely resemble those of bovine neosporosis and ovine toxoplasmosis. Although abortion caused by naturally occurring neosporosis in sheep has not been reported, diagnosticians should carefully distinguish between neosporosis and toxoplasmosis in cases of ovine protozoal abortion unless future investigations exclude the likelihood of naturally acquired neosporosis in sheep.


2015 ◽  
Vol 46 (4) ◽  
pp. 654-659 ◽  
Author(s):  
Maiara Sanitá Tafner Ferreira ◽  
Fernanda Silveira Flores Vogel ◽  
Luis Antonio Sangioni ◽  
Augusto Weber ◽  
Patricia Bräunig ◽  
...  

ABSTRACT: Neosporosis is a parasitic disease caused by the protozoan Neospora caninum which results in major economic losses for cattle breeding due to abortion and other reproductive disorders. Gerbils (Meriones unguiculatus) are commonly used as experimental models for neosporosis due to their high susceptibility to N. caninum infection, both by oocysts ingestion as by tachyzoites/bradyzoites parenteral inoculation. However, the risk of transmission by tachyzoites ingestion is not fully elucidated. In this study, infection of neonate gerbils by N. caninum (NC-1 strain) tachyzoites inoculated by the oral route and the parasite distribution in gerbils' tissues were evaluated by protozoan DNA detection. Seventeen neonate gerbils, aged 4-5 days, were inoculated with 4x105 tachyzoites by the oral route and one gerbil was kept as uninfected control. N. caninum DNA was detected in 100% of the inoculated gerbils, showing that the oral route is effective as a potential route of infection of neonates by N. caninum tachyzoites. N. caninum DNA was reported in all organs evaluated (heart, lungs, kidneys, liver, spleen and brain), with different frequencies. These results showed systemically distributed infection of neonate gerbils after oral inoculation of tachyzoites.


2020 ◽  
Vol 58 (7) ◽  
Author(s):  
Higor Sette Pereira ◽  
Ludmila Tavares e Almeida ◽  
Vitória Fernandes ◽  
Renato Lima Senra ◽  
Patrícia Pereira Fontes ◽  
...  

ABSTRACT Neosporosis has become a concern since it is associated with abortion in cattle. Currently, in situ diagnosis is determined through anamnesis, evaluation of the history, and perception of the clinical signs of the herd. There is no practical and noninvasive test adapted to a large number of samples, which represents a gap for the use of new approaches that provide information about infections and the risks of herds. Here, we performed a search in the Neospora caninum genome by linear B-cell epitopes using immunoinformatic tools aiming to develop a chimeric protein with high potential to bind specifically to antibodies from infected cattle samples. An enzyme-linked immunosorbent assay with the new chimeric antigen was developed and tested with sera from natural field N. caninum-infected bovines. The cross-reactivity of the new antigen was also evaluated using sera from bovines infected by other abortive pathogens, including Trypanosoma vivax, Leptospira sp., Mycobacterium bovis, and Brucella abortus, and enzootic bovine leucosis caused by bovine leukemia virus, as well as with samples of animals infected with Toxoplasma gondii. The assay using the chimeric protein showed 96.6% ± 3.4% of sensitivity in comparison to healthy animal sera. Meanwhile, in relation to false-positive results provided by cross-reactivity with others pathogens, the specificity value was 97.0% ± 2.9%. In conclusion, immunoinformatic tools provide an efficient platform to build an accurate protein to diagnose bovine neosporosis based on serum samples.


Parasitology ◽  
1999 ◽  
Vol 118 (4) ◽  
pp. 363-370 ◽  
Author(s):  
R. ATKINSON ◽  
P. A. W. HARPER ◽  
C. RYCE ◽  
D. A. MORRISON ◽  
J. T. ELLIS

This study compared the biological and genetic properties of a bovine (NC-SweB1) and a canine (NC-Liverpool) isolate of Neospora caninum. A mouse model for CNS infection demonstrated marked differences in pathogenicity between the isolates. NC-Liverpool induced severe clinical signs of neosporosis in 57/58 mice including discoordinated movement, hindlimb paralysis and coat ruffling with severe weight loss. In contrast NC-SweB1 induced similar but less severe symptoms in a much smaller proportion of mice over the same time-period. Statistically significant differences were observed between the isolates in the response (mean weight loss) of mice through time to the different doses inoculated. Histopathological effects on brain tissue reflected the isolate-based differences described above. NC-Liverpool infection resulted in intense inflammatory infiltrates and highly necrotic lesions whereas NC-SweB1 induced a milder meningoencephalitis. Passage in cell-culture over a period of 14 months did not affect the pathogenicity of NC-Liverpool. Immunoblots showed that antibodies to N. caninum appeared earlier in mice inoculated with NC-Liverpool than with NC-SweB1. Finally, RAPD–PCR analysis of NC-Liverpool DNA generated profiles distinct from that observed with DNA from NC-SweB1 or Toxoplasma gondii. In summary this study provides evidence for significant biological and genetic differences between 2 isolates of N. caninum.


Author(s):  
W.L. Steffens ◽  
M.B. Ard ◽  
C.E. Greene ◽  
A. Jaggy

Canine distemper is a multisystemic contagious viral disease having a worldwide distribution, a high mortality rate, and significant central neurologic system (CNS) complications. In its systemic manifestations, it is often presumptively diagnosed on the basis of clinical signs and history. Few definitive antemortem diagnostic tests exist, and most are limited to the detection of viral antigen by immunofluorescence techniques on tissues or cytologic specimens or high immunoglobulin levels in CSF (cerebrospinal fluid). Diagnosis of CNS distemper is often unreliable due to the relatively low cell count in CSF (<50 cells/μl) and the binding of blocking immunoglobulins in CSF to cell surfaces. A more reliable and definitive test might be possible utilizing direct morphologic detection of the etiologic agent. Distemper is the canine equivalent of human measles, in that both involve a closely related member of the Paramyxoviridae, both produce mucosal inflammation, and may produce CNS complications. In humans, diagnosis of measles-induced subacute sclerosing panencephalitis is through negative stain identification of whole or incomplete viral particles in patient CSF.


2021 ◽  
Vol 8 (2) ◽  
pp. 30
Author(s):  
Luis Emilio Fazzio ◽  
Santiago José Raggio ◽  
Juan Facundo Romero ◽  
Juver Membrebe ◽  
Antonio Humberto Hamad Minervino

A safety study on ketoprofen 10% was carried out on pigs using a different dosing and treatment scheme. Forty healthy crossbreed pigs with similar age, weight, and body condition score were distributed into five treatment groups. The pigs were intramuscularly injected once with different doses of ketoprofen: 3 mg/kg (group 1X), 6 mg/kg (group 2X), 9 mg/kg (group 3X). In addition, the 3 mg/kg dosis was administered on three consecutive days (group 1X ext.). Intramuscular injections of saline solution were used in control group (CTL). The pigs were clinically examined throughout the trial and blood samples were taken for hematological and biochemical evaluation on days −4 (before treatment), +3, +7, and +14 (the end of the trial). Any unusual behaviour or clinical signs were reported as potential toxic effects of ketoprofen. Serum measurements showed that none of the ketoprofen doses produced changes in renal or hepatic biochemical parameters, liver enzymes, or total bilirubin. Likewise, hematological assessment indicated no altered parameters or hematocrit percentage in the study groups. These results demonstrate that ketoprofen has no adverse effects in pigs when the doses and scheme evaluated in this study are applied.


2021 ◽  
pp. 104063872199668
Author(s):  
Waléria Borges-Silva ◽  
Mariana M. Rezende-Gondim ◽  
Gideão S. Galvão ◽  
Daniele S. Rocha ◽  
George R. Albuquerque ◽  
...  

Parasites resembling Neospora caninum or Toxoplasma gondii were detected by cytologic examination of cerebrospinal fluid (CSF) from a dog with neurologic disease. The dog became severely ill and was euthanized. Canine tissue homogenates were used for direct parasite isolation in cell culture, bioassay in 2 mouse lineages, and PCR. T. gondii was isolated in monkey kidney cells, and species identity was confirmed by PCR. Inoculated parasites were highly virulent for mice, which developed clinical signs and were euthanized immediately. PCR-RFLP for T. gondii using the cultured isolate (TgDgBA22) was conducted with 12 genetic markers, and a unique recombinant strain was identified. Detection of T. gondii by CSF cytology, although described in humans, had not been reported previously in dogs, to our knowledge, and was crucial for the diagnosis of toxoplasmosis in the examined dog.


2004 ◽  
Vol 46 (4) ◽  
pp. 209-216 ◽  
Author(s):  
Nnamdi Callistus D. Ukwandu ◽  
O. P. G. Nmorsi

Well-structured questionnaire on the perception, impression and response to genitourinary bilharziasis (Genitourinary schistosomiasis) was administered and explained in local languages: 'Igbo' 'Esan' 'Ezon' Itshekiri and Bini to 33815 inhabitants of selected endemic areas in south-eastern Nigeria from January, 1999 to December, 2001. Out of this number, 3815 (11.3%) were properly filled and returned. About 42.0% of the inhabitants admitted knowledge of the disease, while 14 (0.4%) knew about the aetiologic agent. About 181 (5.0%) who responded, admitted procuring treatment, while 100 (5.0%) declined to seek treatment of any sort. The relationships between water-bodies and human activities, and infection were well discussed. Amongst those who admitted knowledge of the disease but no knowledge of its etiologic agent, declined seeking treatment of any kind, but believe the disease is a natural phenomenon in ones developmental stage and therefore of no morbidity and mortality. Laboratory analysis of urine, faeces, semen and HVS was employed to assess questionnaire responses, and in some cases, physical examination was utilized to augment laboratory analysis in confirming urinal diagnosis. Haematuria was only directly related to egg count in the early part of life. Females were significantly haematuric and excreted more ova than males (p < 0.05). Headache (43.0%) and fever (31.0%) were major clinical signs while sexual pains (22.0%) were the least.


Pathogens ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1074
Author(s):  
Natalia Vacani-Martins ◽  
Marcelo Meuser-Batista ◽  
Carina de Lima Pereira dos Santos ◽  
Alejandro Marcel Hasslocher-Moreno ◽  
Andrea Henriques-Pons

Chagas disease was described more than a century ago and, despite great efforts to understand the underlying mechanisms that lead to cardiac and digestive manifestations in chronic patients, much remains to be clarified. The disease is found beyond Latin America, including Japan, the USA, France, Spain, and Australia, and is caused by the protozoan Trypanosoma cruzi. Dr. Carlos Chagas described Chagas disease in 1909 in Brazil, and hepatomegaly was among the clinical signs observed. Currently, hepatomegaly is cited in most papers published which either study acutely infected patients or experimental models, and we know that the parasite can infect multiple cell types in the liver, especially Kupffer cells and dendritic cells. Moreover, liver damage is more pronounced in cases of oral infection, which is mainly found in the Amazon region. However, the importance of liver involvement, including the hepatic immune response, in disease progression does not receive much attention. In this review, we present the very first paper published approaching the liver’s participation in the infection, as well as subsequent papers published in the last century, up to and including our recently published results. We propose that, after infection, activated peripheral T lymphocytes reach the liver and induce a shift to a pro-inflammatory ambient environment. Thus, there is an immunological integration and cooperation between peripheral and hepatic immunity, contributing to disease control.


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