Experimental infection of cats with Cryptosporidium felis

2021 ◽  
pp. 1098612X2110534
Author(s):  
Andrea V Scorza ◽  
Phyllis Tyrrell ◽  
Sara Wennogle ◽  
Ramaswamy Chandrashekar ◽  
Michael R Lappin
Keyword(s):  
Quantitative Pcr ◽  
Experimental Infection ◽  
Fluorescent Antibody ◽  
Clinical Signs ◽  
The Other ◽  
Methylprednisolone Acetate ◽  
Antibody Assay ◽  
Fecal Samples

Objectives The aims of this study were to experimentally inoculate cats with Cryptosporidium felis oocysts and compare fecal detection by fluorescent antibody assay (FA) and quantitative PCR (qPCR), and document clinical signs associated with infection. Methods Cryptosporidium felis oocysts were concentrated from the feces of a naturally infected cat and orally inoculated into six cats that tested negative for C felis by an FA and fecal flotation (FF). Cats were observed daily for the presence of clinical signs consistent with infection. Fecal samples from all cats on days 0 and 9, and one sample per cat (days 18–21), were evaluated by all assays. On day 31, two cats negative for C felis by FF and FA were administered methylprednisolone acetate and all assays were repeated on days 34, 36 and 38. Samples from all cats were tested by FF and FA on days 41, 43, 45 and 48. Results A total of 41 samples were tested, 25 of which were compared by FA and qPCR. Cryptosporidium felis was detected in 2/25 (8%) and in 19/25 (76%) samples by FA and by qPCR, respectively; the other 16 samples were tested by FF and FA. None of the cats was positive for C felis by FF or FA in samples collected on days 0, 9 or 18–21. One, five and six samples tested positive by qPCR on days 0, 9 and 18–21, respectively. The cats administered methylprednisolone acetate tested positive for C felis by FA on day 36 and by qPCR on days 31, 34, 36 and 38. None of the cats showed clinical signs of disease. Conclusions and relevance Clinical signs were not recognized in any of the cats for the duration of the study. FA was insensitive compared with qPCR for detecting cats with subclinical C felis infection.

scholarly journals Rapid Onset of Ulcerative Typhlocolitis in B6.129P2-IL10tm1Cgn (IL-10−/−) Mice Infected with Helicobacter trogontum Is Associated with Decreased Colonization by Altered Schaedler's Flora

2006 ◽  
Vol 74 (12) ◽  
pp. 6615-6623 ◽  
Author(s):  
M. T. Whary ◽  
S. J. Danon ◽  
Y. Feng ◽  
Z. Ge ◽  
N. Sundina ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Quantitative Pcr ◽  
Body Condition ◽  
Experimental Infection ◽  
Clinical Signs ◽  
Interleukin 10 ◽  
Rapid Onset ◽  
Lower Bowel ◽  
Severe Diarrhea ◽  
First Time

ABSTRACT Infection with Helicobacter trogontum, a urease-positive helicobacter isolated from subclinically infected rats, was evaluated in B6.129P2-IL10 tm1Cgn (interleukin-10−/− [IL-10−/−]) and C57BL/6 (B6) mice. In a first experiment, IL-10−/− mice naturally infected with Helicobacter rodentium had subclinical typhlocolitis but developed severe diarrhea and loss of body condition with erosive to ulcerative typhlocolitis within 1 to 3 weeks of experimental infection with H. trogontum. A second experiment demonstrated that helicobacter-free IL-10−/− mice dosed with H. trogontum also developed severe clinical signs and typhlocolitis within 2 to 4 weeks, whereas B6 mice colonized with H. trogontum were resistant to disease. In a third experiment, using helicobacter-free IL-10−/− mice, dosing with H. trogontum resulted in acute morbidity and typhlocolitis within 8 days. Acute typhlocolitis was accompanied by signs of sepsis supported by degenerative hemograms and recovery of Escherichia coli and Proteus spp. from the livers of infected mice. Quantitative PCR data revealed that H. rodentium and H. trogontum may compete for colonization of the lower bowel, as H. trogontum established higher colonization levels in the absence of H. rodentium (P < 0.003). H. trogontum-induced typhlocolitis was also associated with a significant decrease in the levels of colonization by five of eight anaerobes that comprise altered Schaedler's flora (P < 0.002). These results demonstrate for the first time that H. rodentium infection in IL-10−/− mice causes subclinical typhlocolitis and that infection with H. trogontum (with or without H. rodentium) induces a rapid-onset, erosive to ulcerative typhlocolitis which impacts the normal anaerobic flora of the colon and increases the risk of sepsis.

scholarly journals Development of Quantitative PCR Assays Targeting the 16S rRNA Genes of Enterococcus spp. and Their Application to the Identification of Enterococcus Species in Environmental Samples

2012 ◽  
Vol 79 (1) ◽  
pp. 196-204 ◽  
Author(s):  
Hodon Ryu ◽  
Michael Henson ◽  
Michael Elk ◽  
Carlos Toledo-Hernandez ◽  
John Griffith ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Quantitative Pcr ◽  
Water Samples ◽  
The Other ◽  
Rrna Gene ◽  
Fecal Samples ◽  
Content Type ◽  
Specific Assay ◽  
Enterococcal Species

ABSTRACTThe detection of environmental enterococci has been determined primarily by using culture-based techniques that might exclude some enterococcal species as well as those that are nonculturable. To address this, the relative abundances of enterococci were examined by challenging fecal and water samples against a currently available genus-specific assay (Entero1). To determine the diversity of enterococcal species, 16S rRNA gene-based group-specific quantitative PCR (qPCR) assays were developed and evaluated against eight of the most common environmental enterococcal species. Partial 16S rRNA gene sequences of 439 presumptive environmental enterococcal strains were analyzed to study further the diversity of enterococci and to confirm the specificities of group-specific assays. The group-specific qPCR assays showed relatively high amplification rates with targeted species (>98%), although some assays cross-amplified with nontargeted species (1.3 to 6.5%). The results with the group-specific assays also showed that different enterococcal species co-occurred in most fecal samples. The most abundant enterococci in water and fecal samples wereEnterococcus faecalisandEnterococcus faecium, although we identified more water isolates asEnterococcus casseliflavusthan as any of the other species. The prevalence of the Entero1 marker was in agreement with the combined number of positive signals determined by the group-specific assays in most fecal samples, except in gull feces. On the other hand, the number of group-specific assay signals was lower in all water samples tested, suggesting that other enterococcal species are present in these samples. While the results highlight the value of genus- and group-specific assays for detecting the major enterococcal groups in environmental water samples, additional studies are needed to determine further the diversity, distributions, and relative abundances of all enterococcal species found in water.

scholarly journals Epidemiological study of cryptosporidiosis in immunocompetent & immunocompromised children, prepared parasitic antigen and used it in IFAT

2006 ◽  
Vol 30 (1) ◽  
pp. 73-82
Author(s):  
Kawan M. H.
Keyword(s):  
Epidemiological Study ◽  
Experimental Infection ◽  
Fluorescent Antibody ◽  
Clinical Signs ◽  
Watery Diarrhea ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Significant Difference ◽  
Group 2 ◽  
Group 1

An epidemiological study was done on 2 groups of children infectedwith watery diarrhea in some hospitals of Baghdad (group 1: immunocompetentoutpatients, group 2: immunosuppressed inpatients).Modified cold Ziehl Neelsen stain was used in diagnosis. The wholeoocysts antigen prepared after conduction of experimental infection in 2 calvesby using isolated oocysts from children stool, and this prepared antigen wasused in Indirect Fluorescent Antibody Technique (IFAT).The percentage of infection in the 2 groups of outpatients and inpatientswere 21.2% and 32% respectively, and there was a difference in percentage ofinfection between the 3 groups of children, the highest was 36.4% in group withleukemia and received chemical and radiotherapy.There was no significant difference between the two sexes.The clinical signs of experimental infection began after 4 days and continued for10 days. The oocysts appeared in IFAT as spherical in shape with clear wallsand bright yellowish green color. There was no significant difference betweenthe 2 methods used for diagnosis (stain and immunological test).

scholarly journals Some Pathological Study of Experimental Infection of Blastocystis hominis in Mice

2003 ◽  
Vol 27 (1) ◽  
pp. 41-49
Author(s):  
E. H. AL-Taee
Keyword(s):  
Large Intestine ◽  
Experimental Infection ◽  
Clinical Signs ◽  
Inflammatory Cells ◽  
Liver Cells ◽  
The Other ◽  
Laboratory Animals ◽  
Pathological Study ◽  
Blastocystis Hominis ◽  
Stages Of Growth

Blastocystis hominis isolated from patients suffering from diarrhea patients (56) in number visited two hospital in Baghdad were cultured on liver infusion broth to infecting laboratory animals. Albino mice (4-6) wks in age were infected with Blastocystis hominis by inculcation and after the appearance of clinical signs (weakness, diarrhea) the mice were killed and pathological studies were done on organs like intestine, stomach and liver, the lesion showed desquamation of mucosa, severe infiltration of inflammatory cells specially around the parasite, this lesion was more severe in large intestine from the other organs.  The parasite was seen in liver cells and appeared with different stages of growth and replication causing vaculation and necrosis in the hepatocyte.

A Comparison of the Bethesda and New Oxford Methods of Factor VIII Antibody Assay

1982 ◽  
Vol 47 (01) ◽  
pp. 072-075 ◽  
Author(s):  
D E G Austen ◽  
K Lechner ◽  
C R Rizza ◽  
I L Rhymes
Keyword(s):  
Factor Viii ◽  
Excellent Agreement ◽  
International Committee ◽  
The Other ◽  
Antibody Assay ◽  
Collaborative Trial

SummaryA collaborative trial has been carried out under the auspices of the International Committee on Thrombosis and Haemostasis to compare the Bethesda and New Oxford methods of antibody assay. It was found that errors between laboratories were much greater than those within laboratories and each laboratory had a bias whereby it always rated samples high or low with respect to the other laboratories. However there was excellent agreement in the order in which laboratories ranked antibody samples and if a standard antibody sample could be provided there would be a significant improvement in numerical agreement between laboratories. On average, for this exercise, a result for a given sample in Bethesda units was 1.21 times the result in New Oxford units although it must be stressed that this ratio could vary from sample to sample.

Porcine teschovirus, sapelovirus, and enterovirus in Swiss pigs: multiplex RT-PCR investigation of viral frequencies and disease association

2021 ◽  
pp. 104063872110258
Author(s):  
Tamara Stäubli ◽  
Charlotte I. Rickli ◽  
Paul R. Torgerson ◽  
Cornel Fraefel ◽  
Julia Lechmann
Keyword(s):  
Clinical Signs ◽  
Virus Detection ◽  
Common Finding ◽  
Fecal Samples ◽  
High Frequencies ◽  
Reverse Transcription Pcr ◽  
Suckling Piglets ◽  
Porcine Teschovirus ◽  
Fattening Pigs ◽  
Pathology Data

Porcine teschovirus (PTV), sapelovirus (PSV-A), and enterovirus (EV-G) are enteric viruses that can infect pigs and wild boars worldwide. The viruses have been associated with several diseases, primarily gastrointestinal, neurologic, reproductive, and respiratory disorders, but also with subclinical infections. However, for most serotypes, proof of a causal relationship between viral infection and clinical signs is still lacking. In Switzerland, there has been limited investigation of the occurrence of the 3 viruses. We used a modified multiplex reverse-transcription PCR protocol to study the distribution of the viruses in Swiss pigs by testing 363 fecal, brain, and placental or abortion samples from 282 healthy and diseased animals. We did not detect the 3 viruses in 94 placental or abortion samples or in 31 brain samples from healthy pigs. In brain tissue of 81 diseased pigs, we detected 5 PSV-A and 4 EV-G positive samples. In contrast, all 3 viruses were detected at high frequencies in fecal samples of both healthy and diseased pigs. In healthy animals, PTV was detected in 47%, PSV-A in 51%, and EV-G in 70% of the 76 samples; in diseased animals, frequencies in the 81 samples were 54%, 64%, and 68%, respectively. The viruses were detected more frequently in fecal samples from weaned and fattening pigs compared to suckling piglets and sows. Co-detections of all 3 viruses were the most common finding. Based on clinical and pathology data, statistical analysis yielded no evidence for an association of virus detection and disease. Further research is required to determine if pathogenicity is linked to specific serotypes of these viruses.

scholarly journals Experimental infection of indigenous North African goats with goatpox virus

2021 ◽  
Vol 63 (1) ◽  
Author(s):  
Jihane Hamdi ◽  
Zahra Bamouh ◽  
Mohammed Jazouli ◽  
Meryem Alhyane ◽  
Najet Safini ◽  
...  
Keyword(s):  
Antibody Response ◽  
Experimental Infection ◽  
Subcutaneous Tissue ◽  
Severe Disease ◽  
Clinical Signs ◽  
Economic Losses ◽  
North African ◽  
Cutaneous Lesions ◽  
Viral Dna ◽  
Goatpox Virus

Abstract Background Goatpox is a viral disease caused by infection with goatpox virus (GTPV) of the genus Capripoxvirus, Poxviridae family. Capripoxviruses cause serious disease to livestock and contribute to huge economic losses. Goatpox and sheeppox are endemic to Africa, particularly north of the Equator, the Middle East and many parts of Asia. GTPV and sheeppox virus are considered host-specific; however, both strains can cause clinical disease in either goats or sheep with more severe disease in the homologous species and mild or sub-clinical infection in the other. Goatpox has never been reported in Morocco, Algeria or Tunisia despite the huge population of goats living in proximity with sheep in those countries. To evaluate the susceptibility and pathogenicity of indigenous North African goats to GTPV infection, we experimentally inoculated eight locally bred goats with a virulent Vietnamese isolate of GTPV. Two uninfected goats were kept as controls. Clinical examination was carried out daily and blood was sampled for virology and for investigating the antibody response. After necropsy, tissues were collected and assessed for viral DNA using real-time PCR. Results Following the experimental infection, all inoculated goats displayed clinical signs characteristic of goatpox including varying degrees of hyperthermia, loss of appetite, inactivity and cutaneous lesions. The infection severely affected three of the infected animals while moderate to mild disease was noticed in the remaining goats. A high antibody response was developed. High viral DNA loads were detected in skin crusts and nodules, and subcutaneous tissue at the injection site with cycle threshold (Ct) values ranging from 14.6 to 22.9, while lower viral loads were found in liver and lung (Ct = 35.7 and 35.1). The results confirmed subcutaneous tropism of the virus. Conclusion Clinical signs of goatpox were reproduced in indigenous North African goats and confirmed a high susceptibility of the North African goat breed to GTPV infection. A clinical scoring system is proposed that can be applied in GTPV vaccine efficacy studies.

scholarly journals Virus Adaptation Following Experimental Infection of Chickens with a Domestic Duck Low Pathogenic Avian Influenza Isolate from the 2017 USA H7N9 Outbreak Identifies Polymorphic Mutations in Multiple Gene Segments

Viruses ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 1166
Author(s):  
Klaudia Chrzastek ◽  
Karen Segovia ◽  
Mia Torchetti ◽  
Mary Lee Killian ◽  
Mary Pantin-Jackwood ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Experimental Infection ◽  
Clinical Signs ◽  
Interspecies Transmission ◽  
Clinical Samples ◽  
Domestic Duck ◽  
Receptor Binding Site ◽  
Synonymous Mutations ◽  
Field Samples ◽  
Virus Adaptation

In March 2017, highly pathogenic (HP) and low pathogenic (LP) avian influenza virus (AIV) subtype H7N9 were detected from poultry farms and backyard birds in several states in the southeast United States. Because interspecies transmission is a known mechanism for evolution of AIVs, we sought to characterize infection and transmission of a domestic duck-origin H7N9 LPAIV in chickens and genetically compare the viruses replicating in the chickens to the original H7N9 clinical field samples used as inoculum. The results of the experimental infection demonstrated virus replication and transmission in chickens, with overt clinical signs of disease and shedding through both oral and cloacal routes. Unexpectedly, higher levels of virus shedding were observed in some cloacal swabs. Next generation sequencing (NGS) analysis identified numerous non-synonymous mutations at the consensus level in the polymerase genes (i.e., PA, PB1, and PB2) and the hemagglutinin (HA) receptor binding site in viruses recovered from chickens, indicating possible virus adaptation in the new host. For comparison, NGS analysis of clinical samples obtained from duck specimen collected during the outbreak indicated three polymorphic sides in the M1 segment and a minor population of viruses carrying the D139N (21.4%) substitution in the NS1 segment. Interestingly, at consensus level, A/duck/Alabama (H7N9) had isoleucine at position 105 in NP protein, similar to HPAIV (H7N9) but not to LPAIV (H7N9) isolated from the same 2017 influenza outbreak in the US. Taken together, this work demonstrates that the H7N9 viruses could readily jump between avian species, which may have contributed to the evolution of the virus and its spread in the region.

scholarly journals Experimental infection of broiler chicks with Salmonella Typhimurium from pigeon (Columba livia)

2016 ◽  
Vol 37 (4) ◽  
pp. 1919
Author(s):  
Átilla Holanda de Albuquerque ◽  
Régis Siqueira de Castro Teixeira ◽  
Débora Nishi Machado ◽  
Elisângela De Souza Lopes ◽  
Ruben Horn Vasconcelos ◽  
...  
Keyword(s):  
Salmonella Typhimurium ◽  
Clinical Signs ◽  
Columba Livia ◽  
Poultry Feed ◽  
Economic Losses ◽  
Control Group ◽  
Broiler Chicks ◽  
Post Inoculation ◽  
Microbiological Analysis ◽  
Fecal Samples

Several cases of animal and human salmonellosis caused by the Salmonella serotype Typhimurium have been reported. In animals, subclinical infection favors pathogen dissemination through feces. In this context, the domestic pigeon (Columba livia) with an asymptomatic condition may play an important role in the transmission of salmonellosis, through the elimination of contaminated feces in commercial aviaries or in poultry feed facilities, causing economic losses to the poultry industry and presenting a risk to public health. This study aimed to evaluate the mortality, clinical signs and the presence of Salmonella Typhimurium in the feces and organs of chicks previously inoculated with bacteria isolated from a pigeon. One-day-old chicks were distributed in two experimental groups (G1 and G2) of 32 birds each, and a control group of six birds. Two inocula of 0.4 and 0.7 mL with 105 and 106 colony forming units were used in G1 and G2 birds, respectively. At 1, 4, 7 and 14 days post-inoculation (dpi) fecal samples were pooled from each cage and individual cloacal swabs were collected. At 14 dpi, all chicks were euthanized and samples were collected from the liver, spleen, lung, cecum and intestine for microbiological analysis. Mortality was only observed among G2 birds (6.25%). Most birds presented clinical signs of diarrhea at 4 dpi and no symptom as observed at 14 dpi. The results from cloacal swabs demonstrated bacterial elimination in 68.8% and 53.1% of G2 and G1 birds, respectively at 1 dpi. Additionally, fecal samples had elevated bacterial shedding in all four periods of observation , with a higher excretion at 4 dpi (62.5%) for both groups. Among G2 birds, 74.2% were positive for the pathogen in the intestine; G1 birds presented the lowest rate of lung infection (29%), and both groups had more than 50% positivity for liver and caeca. The results revealed that infected chicks with a Salmonella Typhimurium strains isolated from pigeons may host the pathogen in several organs, and simultaneously present diarrheic disorders with significant levels of bacterial excretion in feces.

The influence of dietary carbohydrates on experimental infection withTrichuris suisin pigs

Parasitology ◽  
2005 ◽  
Vol 132 (6) ◽  
pp. 857-865 ◽  
Author(s):  
L. E. THOMSEN ◽  
S. PETKEVIČIUS ◽  
K. E. BACH KNUDSEN ◽  
A. ROEPSTORFF
Keyword(s):  
Sugar Beet ◽  
Experimental Infection ◽  
Diet Group ◽  
The Other ◽  
Post Inoculation ◽  
Dietary Carbohydrates ◽  
Barley Flour ◽  
Faecal Egg Counts ◽  
The Difference ◽  
Fermentable Carbohydrates

Two experiments (Exps 1 and 2) were carried out to study the effect of dietary carbohydrates on the establishment ofTrichuris suisin pigs. Two experimental diets based on barley flour were used; Diet 1 was supplemented with non-fermentable carbohydrates from oat hull meal, while Diet 2 was supplemented with fermentable carbohydrates from sugar beet fibre and inulin. In Exp. 1, thirty-two pigs were allocated randomly into 4 groups. Two groups were fed Diet 1 and 2 groups were fed Diet 2. Pigs from one of each diet group were inoculated with 2000 infectiveT. suiseggs each and the other two groups were uninfected controls. All pigs were slaughtered 8 weeks post-inoculation (p.i.). In Exp. 2, twenty-four pigs were allocated randomly into 2 groups and fed Diet 1 or Diet 2, respectively. All the pigs were inoculated with 2000 infectiveT. suiseggs. Six pigs from each group were slaughtered 8 weeks p.i. and the remaining 6 pigs from each group were slaughtered 12 weeks p.i. Infections were followed by faecal egg counts and worm burdens were assessed at necropsy. Pigs fed Diet 2 had lower egg counts in both experiments; in Exp. 2 the difference was significant (P<0·05). No differences were found in worm burdens 8 weeks p.i. in both experiments, however, worms from pigs on Diet 2 were significantly shorter (P<0·0001). Pigs fed Diet 2 and slaughtered 12 weeks p.i. had significantly lower worm counts (P<0·01) compared to pigs fed Diet 1. The results indicate that fermentable carbohydrates do not affect the establishment ofT. suisin naïve pigs, but result in earlier expulsion and reduced growth of the established worms. Thus, diets with highly fermentable carbohydrates may be used in the control ofT. suis.

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