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1.
Appl Environ Microbiol ; 80(5): 1639-44, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24375131

RESUMO

Recent studies have suggested a potential role for wild birds in zoonotic transmission of Campylobacter jejuni, the leading cause of gastroenteritis in humans worldwide. In this study, we detected Campylobacter spp. in 66.9% (85/127) of free-ranging American crows (Corvus brachyrhyncos) sampled in the Sacramento Valley of California in 2012 and 2013. Biochemical testing and sequence analysis of 16S rRNA revealed that 93% of isolates (n = 70) were C. jejuni, with cytolethal distending toxin (CDT) and flagellin A genes detected by PCR in 20% and 46% of the C. jejuni isolates (n = 59), respectively. The high prevalence of C. jejuni, coupled with the occurrence of known virulence markers CDT and flagellin A, demonstrates that crows shed Campylobacter spp. in their feces that are potentially pathogenic to humans. Crows are abundant in urban, suburban, and agricultural settings, and thus further study to determine their role in zoonotic transmission of Campylobacter will inform public health.


Assuntos
Doenças das Aves/epidemiologia , Doenças das Aves/microbiologia , Infecções por Campylobacter/veterinária , Campylobacter jejuni/isolamento & purificação , Corvos/microbiologia , Animais , Toxinas Bacterianas/genética , Técnicas de Tipagem Bacteriana , California , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/microbiologia , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Flagelina/genética , Humanos , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Prevalência , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
2.
Appl Environ Microbiol ; 79(6): 1859-65, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23315738

RESUMO

Constructed wetland systems are used to reduce pollutants and pathogens in wastewater effluent, but comparatively little is known about pathogen transport through natural wetland habitats. Fecal protozoans, including Cryptosporidium parvum, Giardia lamblia, and Toxoplasma gondii, are waterborne pathogens of humans and animals, which are carried by surface waters from land-based sources into coastal waters. This study evaluated key factors of coastal wetlands for the reduction of protozoal parasites in surface waters using settling column and recirculating mesocosm tank experiments. Settling column experiments evaluated the effects of salinity, temperature, and water type ("pure" versus "environmental") on the vertical settling velocities of C. parvum, G. lamblia, and T. gondii surrogates, with salinity and water type found to significantly affect settling of the parasites. The mesocosm tank experiments evaluated the effects of salinity, flow rate, and vegetation parameters on parasite and surrogate counts, with increased salinity and the presence of vegetation found to be significant factors for removal of parasites in a unidirectional transport wetland system. Overall, this study highlights the importance of water type, salinity, and vegetation parameters for pathogen transport within wetland systems, with implications for wetland management, restoration efforts, and coastal water quality.


Assuntos
Cryptosporidium parvum/isolamento & purificação , Giardia lamblia/isolamento & purificação , Toxoplasma/isolamento & purificação , Purificação da Água/métodos , Água/parasitologia , Áreas Alagadas , Animais , Humanos , Concentração de Íons de Hidrogênio , Microesferas , Salinidade , Temperatura , Água/química
3.
Microb Ecol ; 65(4): 982-94, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23392641

RESUMO

Given their coastal site fidelity and opportunistic foraging behavior, harbor seals (Phoca vitulina) may serve as sentinels for coastal ecosystem health. Seals using urbanized coastal habitat can acquire enteric bacteria, including Vibrio that may affect their health. To understand Vibrio dynamics in seals, demographic and environmental factors were tested for predicting potentially virulent Vibrio in free-ranging and stranded Pacific harbor seals (Phoca vitulina richardii) off California. Vibrio prevalence did not vary with season and was greater in free-ranging seals (29 %, n = 319) compared with stranded seals (17 %, n = 189). Of the factors tested, location, turbidity, and/or salinity best predicted Vibrio prevalence in free-ranging seals. The relationship of environmental factors with Vibrio prevalence differed by location and may be related to oceanographic or terrestrial contributions to water quality. Vibrio parahaemolyticus, Vibrio alginolyticus, and Vibrio cholerae were observed in seals, with V. cholerae found almost exclusively in stranded pups and yearlings. Additionally, virulence genes (trh and tdh) were detected in V. parahaemolyticus isolates. Vibrio cholerae isolates lacked targeted virulence genes, but were hemolytic. Three out of four stranded pups with V. parahaemolyticus (trh+ and/or tdh+) died in rehabilitation, but the role of Vibrio in causing mortality is unclear, and Vibrio expression of virulence genes should be investigated. Considering that humans share the environment and food resources with seals, potentially virulent Vibrio observed in seals also may be of concern to human health.


Assuntos
Proteínas de Bactérias/genética , Phoca/microbiologia , Vibrioses/veterinária , Vibrio/isolamento & purificação , Fatores de Virulência/genética , Animais , Animais Selvagens/microbiologia , Proteínas de Bactérias/metabolismo , California , Humanos , Mamíferos/microbiologia , Vibrio/classificação , Vibrio/genética , Vibrioses/microbiologia , Fatores de Virulência/metabolismo
4.
Microb Ecol ; 65(4): 928-33, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23250114

RESUMO

Aquatic macroaggregates (flocs ≥ 0.5 mm) provide an important mechanism for vertical flux of nutrients and organic matter in aquatic ecosystems, yet their role in the transport and fate of zoonotic pathogens is largely unknown. Terrestrial pathogens that enter coastal waters through contaminated freshwater runoff may be especially prone to flocculation due to fluid dynamics and electrochemical changes that occur where fresh and marine waters mix. In this study, laboratory experiments were conducted to evaluate whether zoonotic pathogens (Cryptosporidium, Giardia, Salmonella) and a virus surrogate (PP7) are associated with aquatic macroaggregates and whether pathogen aggregation is enhanced in saline waters. Targeted microorganisms showed increased association with macroaggregates in estuarine and marine waters, as compared with an ultrapure water control and natural freshwater. Enrichment factor estimations demonstrated that pathogens are 2-4 orders of magnitude more concentrated in aggregates than in the estuarine and marine water surrounding the aggregates. Pathogen incorporation into aquatic macroaggregates may influence their transmission to susceptible hosts through settling and subsequent accumulation in zones where aggregation is greatest, as well as via enhanced uptake by invertebrates that serve as prey for marine animals or as seafood for humans.


Assuntos
Bactérias/isolamento & purificação , Bacteriófagos/isolamento & purificação , Água Doce/parasitologia , Giardia/isolamento & purificação , Água do Mar/parasitologia , Microbiologia da Água , Zoonoses/parasitologia , Animais , Bactérias/genética , Bacteriófagos/genética , Ecossistema , Água Doce/microbiologia , Água Doce/virologia , Giardia/genética , Água do Mar/microbiologia , Água do Mar/virologia , Zoonoses/microbiologia , Zoonoses/transmissão , Zoonoses/virologia
5.
J Zoo Wildl Med ; 44(2): 376-88, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23805556

RESUMO

Blood analytes are critical for evaluating the general health of cetacean populations, so it is important to understand the intrinsic variability of hematology and serum chemistry values. Previous studies have reported data for follow-up periods of several years in managed and wild populations, but studies over long periods of time (> 20 yr) have not been reported. The study objective was to identify the influences of partitioning characteristics on hematology and serum chemistry analytes of apparently healthy managed beluga (Delphinapterus leucas). Blood values from 31 managed belugas, at three facilities, collected over 22 yr, were assessed for seasonal variation and aging trends, and evaluated for biologic variation among and within individuals. Linear mixed effects models assessed the relationship between the analytes and sex, age, season, facility location, ambient air temperature, and photoperiod. Sex differences in analytes and associations with increasing age were observed. Seasonal variation was observed for hemoglobin, hematocrit, mean corpuscular volume, monocytes, alkaline phosphatase, total bilirubin, cholesterol, and triglycerides. Facilities were associated with larger effects on analyte values compared to other covariates, whereas age, sex, and ambient temperature had smaller effects compared to facility and season. Present findings provide important baseline information for future health monitoring efforts. Interpretation of blood analytes and animal health in managed and wild populations over time is aided by having available typical levels for the species and reference intervals for the degree to which individual animals vary from the species average and from their own baseline levels during long-term monitoring.


Assuntos
Beluga/sangue , Testes Hematológicos/veterinária , Animais , Coleta de Amostras Sanguíneas , Feminino , Masculino , Análise Multivariada , Valores de Referência , Fatores Sexuais , Fatores de Tempo
6.
J Clin Microbiol ; 50(12): 4131-3, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23052307

RESUMO

Pulsed-field gel electrophoresis (PFGE) was used to type 128 Streptococcus infantarius subsp. coli isolates from sea otters and mussels. Six SmaI PFGE groups were detected, with one predominant group representing 57% of the isolates collected over a wide geographic region. Several sea otter and mussel isolates were highly related, suggesting that an environmental infection source is possible.


Assuntos
Endocardite/veterinária , Lontras/microbiologia , Sepse/veterinária , Infecções Estreptocócicas/veterinária , Streptococcus/classificação , Streptococcus/isolamento & purificação , Animais , Bivalves/microbiologia , Análise por Conglomerados , Eletroforese em Gel de Campo Pulsado , Endocardite/microbiologia , Genótipo , Epidemiologia Molecular , Tipagem Molecular , Sepse/microbiologia , Infecções Estreptocócicas/microbiologia , Streptococcus/genética
7.
Appl Environ Microbiol ; 78(24): 8762-72, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23042185

RESUMO

The risk of disease transmission from waterborne protozoa is often dependent on the origin (e.g., domestic animals versus wildlife), overall parasite load in contaminated waterways, and parasite genotype, with infections being linked to runoff or direct deposition of domestic animal and wildlife feces. Fecal samples collected from domestic animals and wildlife along the central California coast were screened to (i) compare the prevalence and associated risk factors for fecal shedding of Cryptosporidium and Giardia species parasites, (ii) evaluate the relative importance of animal host groups that contribute to pathogen loading in coastal ecosystems, and (iii) characterize zoonotic and host-specific genotypes. Overall, 6% of fecal samples tested during 2007 to 2010 were positive for Cryptosporidium oocysts and 15% were positive for Giardia cysts. Animal host group and age class were significantly associated with detection of Cryptosporidium and Giardia parasites in animal feces. Fecal loading analysis revealed that infected beef cattle potentially contribute the greatest parasite load relative to other host groups, followed by wild canids. Beef cattle, however, shed host-specific, minimally zoonotic Cryptosporidium and Giardia duodenalis genotypes, whereas wild canids shed potentially zoonotic genotypes, including G. duodenalis assemblages A and B. Given that the parasite genotypes detected in cattle were not zoonotic, the public health risk posed by protozoan parasite shedding in cattle feces may be lower than that posed by other animals, such as wild canids, that routinely shed zoonotic genotypes.


Assuntos
Criptosporidiose/veterinária , Cryptosporidium/classificação , Cryptosporidium/genética , Giardia lamblia/classificação , Giardia lamblia/genética , Giardíase/veterinária , Animais , Animais Domésticos , Animais Selvagens , California/epidemiologia , Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , DNA de Protozoário/química , DNA de Protozoário/genética , Fezes/parasitologia , Genótipo , Giardia lamblia/isolamento & purificação , Giardíase/parasitologia , Dados de Sequência Molecular , Prevalência , Análise de Sequência de DNA
8.
Appl Environ Microbiol ; 78(10): 3606-13, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22427504

RESUMO

Fecal pathogen contamination of watersheds worldwide is increasingly recognized, and natural wetlands may have an important role in mitigating fecal pathogen pollution flowing downstream. Given that waterborne protozoa, such as Cryptosporidium and Giardia, are transported within surface waters, this study evaluated associations between fecal protozoa and various wetland-specific and environmental risk factors. This study focused on three distinct coastal California wetlands: (i) a tidally influenced slough bordered by urban and agricultural areas, (ii) a seasonal wetland adjacent to a dairy, and (iii) a constructed wetland that receives agricultural runoff. Wetland type, seasonality, rainfall, and various water quality parameters were evaluated using longitudinal Poisson regression to model effects on concentrations of protozoa and indicator bacteria (Escherichia coli and total coliform). Among wetland types, the dairy wetland exhibited the highest protozoal and bacterial concentrations, and despite significant reductions in microbe concentrations, the wetland could still be seen to influence water quality in the downstream tidal wetland. Additionally, recent rainfall events were associated with higher protozoal and bacterial counts in wetland water samples across all wetland types. Notably, detection of E. coli concentrations greater than a 400 most probable number (MPN) per 100 ml was associated with higher Cryptosporidium oocyst and Giardia cyst concentrations. These findings show that natural wetlands draining agricultural and livestock operation runoff into human-utilized waterways should be considered potential sources of pathogens and that wetlands can be instrumental in reducing pathogen loads to downstream waters.


Assuntos
Cryptosporidium/isolamento & purificação , Enterobacteriaceae/isolamento & purificação , Giardia/isolamento & purificação , Microbiologia da Água , Água/parasitologia , Áreas Alagadas , Carga Bacteriana , California/epidemiologia , Escherichia coli , Humanos , Carga Parasitária , Estações do Ano , Tempo (Meteorologia)
9.
J Am Vet Med Assoc ; 238(6): 773-83, 2011 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-21401435

RESUMO

OBJECTIVE: To determine types and estimate prevalence of potentially zoonotic enteric pathogens shed by wild animals admitted to either of 2 wildlife hospitals and to characterize distribution of these pathogens and of aerobic bacteria in a hospital environment. DESIGN: Cross-sectional study. SAMPLE: Fecal samples from 338 animals in 2 wildlife hospitals and environmental samples from 1 wildlife hospital. PROCEDURES: Fecal samples were collected within 24 hours of hospital admission. Environmental samples were collected from air and surfaces. Samples were tested for zoonotic pathogens via culture techniques and biochemical analyses. Prevalence of pathogen shedding was compared among species groups, ages, sexes, and seasons. Bacterial counts were determined for environmental samples. RESULTS: Campylobacter spp, Vibrio spp, Salmonella spp, Giardia spp, and Cryptosporidium spp (alone or in combination) were detected in 105 of 338 (31%) fecal samples. Campylobacter spp were isolated only from birds. Juvenile passerines were more likely to shed Campylobacter spp than were adults; prevalence increased among juvenile passerines during summer. Non-O1 serotypes of Vibrio cholerae were isolated from birds; during an oil-spill response, 9 of 10 seabirds screened were shedding this pathogen, which was also detected in environmental samples. Salmonella spp and Giardia spp were isolated from birds and mammals; Cryptosporidium spp were isolated from mammals only. Floors of animal rooms had higher bacterial counts than did floors with only human traffic. CONCLUSIONS AND CLINICAL RELEVANCE: Potentially zoonotic enteric pathogens were identified in samples from several species admitted to wildlife hospitals, indicating potential for transmission if prevention is not practiced.


Assuntos
Animais Selvagens , Bactérias/isolamento & purificação , Aves/microbiologia , Hospitais Veterinários , Mamíferos/microbiologia , Zoonoses/microbiologia , Animais , Bactérias/classificação , California/epidemiologia , Estudos Transversais , Fezes/microbiologia , Humanos , Zoonoses/epidemiologia
10.
Appl Environ Microbiol ; 76(20): 6821-8, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20802072

RESUMO

The flux of terrestrially derived pathogens to coastal waters presents a significant health risk to marine wildlife, as well as to humans who utilize the nearshore for recreation and seafood harvest. Anthropogenic changes in natural habitats may result in increased transmission of zoonotic pathogens to coastal waters. The objective of our work was to evaluate how human-caused alterations of coastal landscapes in California affect the transport of Toxoplasma gondii to estuarine waters. Toxoplasma gondii is a protozoan parasite that is excreted in the feces of infected felids and is thought to reach coastal waters in contaminated runoff. This zoonotic pathogen causes waterborne toxoplasmosis in humans and is a significant cause of death in threatened California sea otters. Surrogate particles that mimic the behavior of T. gondii oocysts in water were released in transport studies to evaluate if the loss of estuarine wetlands is contributing to an increased flux of oocysts into coastal waters. Compared to vegetated sites, more surrogates were recovered from unvegetated mudflat habitats, which represent degraded wetlands. Specifically, in Elkhorn Slough, where a large proportion of otters are infected with T. gondii, erosion of 36% of vegetated wetlands to mudflats may increase the flux of oocysts by more than 2 orders of magnitude. Total degradation of wetlands may result in increased Toxoplasma transport of 6 orders of magnitude or more. Destruction of wetland habitats along central coastal California may thus facilitate pathogen pollution in coastal waters with detrimental health impacts to wildlife and humans.


Assuntos
Parasitologia/métodos , Água do Mar/microbiologia , Microbiologia do Solo , Toxoplasma/isolamento & purificação , Áreas Alagadas , California , Humanos , Oocistos
11.
Appl Environ Microbiol ; 76(17): 5802-14, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20639358

RESUMO

The value of Bacteroidales genetic markers and fecal indicator bacteria (FIB) to predict the occurrence of waterborne pathogens was evaluated in ambient waters along the central California coast. Bacteroidales host-specific quantitative PCR (qPCR) was used to quantify fecal bacteria in water and provide insights into contributing host fecal sources. Over 140 surface water samples from 10 major rivers and estuaries within the Monterey Bay region were tested over 14 months with four Bacteroidales-specific assays (universal, human, dog, and cow), three FIB (total coliforms, fecal coliforms, and enterococci), two protozoal pathogens (Cryptosporidium and Giardia spp.), and four bacterial pathogens (Campylobacter spp., Escherichia coli O157:H7, Salmonella spp., and Vibrio spp.). Indicator and pathogen distribution was widespread, and detection was not highly seasonal. Vibrio cholerae was detected most frequently, followed by Giardia, Cryptosporidium, Salmonella, and Campylobacter spp. Bayesian conditional probability analysis was used to characterize the Bacteroidales performance assays, and the ratios of concentrations determined using host-specific and universal assays were used to show that fecal contamination from human sources was more common than livestock or dog sources in coastal study sites. Correlations were seen between some, but not all, indicator-pathogen combinations. The ability to predict pathogen occurrence in relation to indicator threshold cutoff levels was evaluated using a weighted measure that showed the universal Bacteroidales genetic marker to have a comparable or higher mean predictive potential than standard FIB. This predictive ability, in addition to the Bacteroidales assays providing information on contributing host fecal sources, supports using Bacteroidales assays in water quality monitoring programs.


Assuntos
Bacteroidetes/genética , Rios/microbiologia , Rios/parasitologia , Água do Mar/microbiologia , Água do Mar/parasitologia , Animais , Carga Bacteriana/métodos , Bacteroidetes/isolamento & purificação , California , Bovinos , Cryptosporidium/isolamento & purificação , Cães , Enterobacteriaceae/isolamento & purificação , Enterococcus/isolamento & purificação , Monitoramento Ambiental/métodos , Giardia/isolamento & purificação , Humanos , Reação em Cadeia da Polimerase/métodos , Estatística como Assunto
12.
J Environ Qual ; 39(5): 1782-9, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-21043283

RESUMO

A survey of storm runoff fecal coliform bacteria (FCB) from working farm and ranch pastures is presented in conjunction with a survey of FCB in manure management systems (MMS). The cross-sectional survey of pasture runoff was conducted on 34 pastures on five different dairies over 2 yr under varying conditions of precipitation, slope, manure management, and use of conservation practices such as vegetative filter strips. The MMS cross-sectional survey consisted of samples collected during 1 yr on nine different dairies from six loafing barns, nine primary lagoons, 12 secondary lagoons, and six irrigation sample points. Pasture runoff samples were additionally analyzed for Cryptosporidium sp. and Giardia duodenalis, whereby detectable concentrations occurred sporadically at higher FCB concentrations resulting in poor correlations with FCB. Prevalence of both parasites was lower relative to high-use areas studied simultaneously on these same farms. Application of manure to pastures more than 2 wk in advance of storm-associated runoff was related to a > or =80% reduction in FCB concentration and load compared to applications within 2 wk before a runoff event. For every 10 m of buffer length, a 24% reduction in FCB concentration was documented. A one-half (75%), one (90%), and two (99%) log10 reduction in manure FCB concentration was observed for manure holding times in MMS of approximately 20, 66, and 133 d, respectively. These results suggest that there are several management and conservation practices for working farms that may result in reduced FCB fluxes from agricultural operations.


Assuntos
Indústria de Laticínios , Microbiologia da Água , Animais , California , Esterco
13.
BMC Infect Dis ; 9: 184, 2009 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-19930709

RESUMO

BACKGROUND: Shigellosis causes diarrheal disease in humans from both developed and developing countries, and multi-drug resistance is an emerging problem. The objective of this study is to present a unified approach that can be used to characterize endemic and outbreak patterns of shigellosis using use a suite of epidemiologic and molecular techniques. The approach is applied to a California case study example of endemic shigellosis at the population level. METHODS: Epidemiologic patterns were evaluated with respect to demographics, multi-drug resistance, antimicrobial resistance genes, plasmid profiles, and pulsed-field gel electrophoresis (PFGE) fingerprints for the 43 Shigella isolates obtained by the Monterey region health departments over the two year period from 2004-2005. RESULTS: The traditional epidemiologic as well as molecular epidemiologic findings were consistent with endemic as compared to outbreak shigellosis in this population. A steady low level of cases was observed throughout the study period and high diversity was observed among strains. In contrast to most studies in developed countries, the predominant species was Shigella flexneri (51%) followed closely by S. sonnei (49%). Over 95% of Shigella isolates were fully resistant to three or more antimicrobial drug subclasses, and 38% of isolates were resistant to five or more subclasses. More than half of Shigella strains tested carried the tetB, catA, or blaTEM genes for antimicrobial resistance to tetracycline, chloramphenicol, and ampicillin, respectively. CONCLUSION: This study shows how epidemiologic patterns at the host and bacterial population levels can be used to investigate endemic as compared to outbreak patterns of shigellosis in a community. Information gathered as part of such investigations will be instrumental in identifying emerging antimicrobial resistance, for developing treatment guidelines appropriate for that community, and to provide baseline data with which to compare outbreak strains in the future.


Assuntos
Surtos de Doenças , Disenteria Bacilar/epidemiologia , Epidemiologia Molecular/métodos , Shigella/classificação , Adolescente , Distribuição por Idade , California/epidemiologia , Criança , Pré-Escolar , DNA Bacteriano/genética , Farmacorresistência Bacteriana Múltipla/genética , Disenteria Bacilar/microbiologia , Eletroforese em Gel de Campo Pulsado , Humanos , Masculino , Testes de Sensibilidade Microbiana , Plasmídeos , Reação em Cadeia da Polimerase , Shigella/genética , Shigella/isolamento & purificação
14.
J Microbiol Methods ; 65(3): 367-79, 2006 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-16181691

RESUMO

Bivalve molluscs concentrate Cryptosporidium oocysts from fecal-contaminated aquatic environments and are therefore useful in monitoring water quality. A real-time TaqMan polymerase chain reaction (PCR) system was developed to allow for large scale quantitative detection of Cryptosporidium spp. in mussels (Mytilus californianus). The TaqMan sensitivity and specificity were compared to conventional PCR and direct immunofluorescent antibody (DFA) assays, with and without immunomagnetic separation (IMS), to identify the best method for parasite detection in mussel hemolymph, gill washings and digestive glands. TaqMan PCR and two conventional PCR systems all detected 1 or more oocysts spiked into 1 ml hemolymph samples. The minimum oocyst detection limit in spiked 5 ml gill wash and 1 g digestive gland samples tested by TaqMan PCR and DFA was 100 oocysts, with a 1 log(10) improvement when samples were first processed by IMS. For tank exposed mussels, TaqMan and conventional PCR methods detected C. parvum in <5% of hemolymph samples. No gill washings from these same mussels tested positive by TaqMan PCR or DFA analysis even with IMS concentration. All methods detected the highest prevalence of C. parvum-positive samples in digestive gland tissues of exposed mussels. In conclusion, the most sensitive method for the detection of C. parvum in oocyst-exposed mussels was IMS concentration with DFA detection: 80% of individual and 100% of pooled digestive gland samples tested positive. TaqMan PCR was comparable to conventional PCR for detection of C. parvum oocysts in mussels and additionally allowed for automated testing, high throughput, and semi-quantitative results.


Assuntos
Cryptosporidium/isolamento & purificação , Mytilus/parasitologia , Animais , Cryptosporidium/classificação , Cryptosporidium/genética , Cryptosporidium/crescimento & desenvolvimento , DNA de Protozoário/análise , Técnica Direta de Fluorescência para Anticorpo , Separação Imunomagnética , Oocistos/genética , Oocistos/isolamento & purificação , Reação em Cadeia da Polimerase , Sensibilidade e Especificidade , Taq Polimerase
15.
Int J Parasitol ; 35(6): 673-84, 2005 May.
Artigo em Inglês | MEDLINE | ID: mdl-15862580

RESUMO

This study evaluated clams as bioindicators of fecal protozoan contamination using three approaches: (i) clam tissue spiking experiments to compare several detection techniques; (ii) clam tank exposure experiments to evaluate clams that had filtered Cryptosporidium oocysts from inoculated water under a range of simulated environmental conditions; (iii) sentinel clam outplanting to assess the distribution and magnitude of fecal contamination in three riverine systems in California. Our spiking and tank experiments showed that direct fluorescent antibody (DFA), immunomagnetic separation (IMS) in combination with DFA, and PCR techniques could be used to detect Cryptosporidium in clam tissues. The most analytically sensitive technique was IMS concentration with DFA detection of oocysts in clam digestive gland tissues, which detected 10 oocysts spiked into a clam digestive gland 83% of the time. In the tank experiment, oocyst dose and clam collection time were significant predictors for detecting Cryptosporidium parvum oocysts in clams. In the wild clam study, Cryptosporidium and Giardia were detected in clams from all three study regions by IMS-DFA analysis of clam digestive glands, with significant variation by sampling year and season. The presence of C. parvum DNA in clams from riverine ecosystems was confirmed with PCR and DNA sequence analysis.


Assuntos
Bivalves/parasitologia , Cryptosporidium/isolamento & purificação , Fezes/parasitologia , Água Doce/parasitologia , Giardia/isolamento & purificação , Poluição da Água , Animais , Sequência de Bases , Biomarcadores/análise , California , Cryptosporidium parvum/genética , Cryptosporidium parvum/isolamento & purificação , DNA de Protozoário/análise , Ecossistema , Técnica Direta de Fluorescência para Anticorpo/métodos , Separação Imunomagnética/métodos , Dados de Sequência Molecular , Oocistos/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Fatores de Risco
16.
J Wildl Dis ; 50(1): 140-2, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24171565

RESUMO

A real-time PCR protocol for detecting Mycobacterium bovis in feces was evaluated in bovine tuberculosis-infected African buffalo (Syncerus caffer). Fecal samples spiked with 1.42 × 10(3) cells of M. bovis culture/g and Bacille Calmette-Guérin standards with 1.58 × 10(1) genome copies/well were positive by real-time PCR but all field samples were negative.


Assuntos
Búfalos/microbiologia , Fezes/microbiologia , Mycobacterium bovis/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Tuberculose/veterinária , Animais , Reação em Cadeia da Polimerase em Tempo Real/métodos , Tuberculose/diagnóstico , Tuberculose/epidemiologia
17.
J Wildl Dis ; 50(1): 21-30, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24171566

RESUMO

Mountain gorillas (Gorilla beringei beringei) are critically endangered primates surviving in two isolated populations in protected areas within the Virunga Massif of Rwanda, Uganda, the Democratic Republic of Congo, and in Bwindi Impenetrable National Park in Uganda. Mountain gorillas face intense ecologic pressures due to their proximity to humans. Human communities outside the national parks, and numerous human activities within the national parks (including research, tourism, illegal hunting, and anti-poaching patrols), lead to a high degree of contact between mountain gorillas and wildlife, domestic animals, and humans. To assess the pathogen transmission potential between wildlife and livestock, feces of mountain gorillas, forest buffalo (Syncerus caffer nanus), and domestic cattle (Bos taurus) in Rwanda were examined for the parasites Giardia and Cryptosporidium. Giardia was found in 9% of mountain gorillas, 6% of cattle, and 2% of forest buffalo. Our study represents the first report of Giardia prevalence in forest buffalo. Cryptosporidium-like particles were also observed in all three species. Molecular characterization of Giardia isolates identified zoonotic genotype assemblage B in the gorilla samples and assemblage E in the cattle samples. Significant spatial clustering of Giardia-positive samples was observed in one sector of the park. Although we did not find evidence for transmission of protozoa from forest buffalo to mountain gorillas, the genotypes of Giardia samples isolated from gorillas have been reported in humans, suggesting that the importance of humans in this ecosystem should be more closely evaluated.


Assuntos
Doenças dos Símios Antropoides/epidemiologia , Búfalos/parasitologia , Doenças dos Bovinos/epidemiologia , Giardíase/veterinária , Gorilla gorilla , Animais , Doenças dos Símios Antropoides/parasitologia , Doenças dos Símios Antropoides/transmissão , Bovinos , Doenças dos Bovinos/parasitologia , Doenças dos Bovinos/transmissão , Reservatórios de Doenças/parasitologia , Reservatórios de Doenças/veterinária , Fezes/parasitologia , Giardia/isolamento & purificação , Giardíase/epidemiologia , Giardíase/parasitologia , Giardíase/transmissão , Gorilla gorilla/parasitologia , Humanos , Contagem de Ovos de Parasitas/veterinária , Ruanda/epidemiologia , Zoonoses
18.
Infect Genet Evol ; 24: 146-56, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24632451

RESUMO

The Pacific Coast band-tailed pigeon (Patagioenas fasciata monilis) is a migratory game bird of North America that is at risk for population decline. Epidemics of avian trichomonosis caused by upper digestive tract infection with Trichomonas spp. protozoa in these and other doves and pigeons of the United States are sporadic, but can involve tens of thousands of birds in a single event. Herein, we analyze the role of trichomonosis in band-tailed pigeon mortality and relate spatial, temporal and demographic patterns of parasite transmission to the genetic background of the infecting organism. Infections were most common in adult birds and prevalence was high in band-tailed pigeons sampled at mortality events (96%) and rehabilitation centers (36%) compared to those that were hunter-killed (11%) or live-caught (4%). During non-epidemic periods, animals were primarily infected with T. gallinae Fe-hydrogenase subtype A2, and were less often infected with either T. gallinae subtype A1 (the British finch epidemic strain), T. stableri n. sp. (a T. vaginalis-like species), or Tritrichomonas blagburni n. sp.-like organisms. Birds sampled during multiple epidemics in California were only infected with T. gallinae subtype A2 and T. stableri. The non-clonal etiology of avian trichomonosis outbreaks in band-tailed pigeons and the risk of spill-over to raptor and passerine species highlights the need for additional studies that clarify the host range and evolutionary relationships between strains of Trichomonas spp. in regions of trichomonosis endemicity.


Assuntos
Columbidae/parasitologia , Tentilhões/parasitologia , Tricomoníase/epidemiologia , Tricomoníase/veterinária , Trichomonas/genética , Animais , Doenças das Aves/parasitologia , California/epidemiologia , DNA de Protozoário/genética , Especificidade de Hospedeiro , Dados de Sequência Molecular , Trichomonas/classificação , Tricomoníase/mortalidade , Tricomoníase/transmissão
19.
Environ Sci Process Impacts ; 15(5): 1041-51, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23552731

RESUMO

Fecal pathogens are transported from a variety of sources in multi-use ecosystems such as upper Cook Inlet (CI), Alaska, which includes the state's urban center and is highly utilized by humans and animals. This study used a novel water quality testing approach to evaluate the presence and host sources of potential fecal pathogens in surface waters and sediments from aquatic ecosystems in upper CI. Matched water and sediment samples, along with effluent from a municipal wastewater treatment facility, were screened for Salmonella spp., Vibrio spp., Cryptosporidium spp., Giardia spp., and noroviruses. Additionally, Bacteroidales spp. for microbial source tracking, and the fecal indicator bacteria Enterococcus spp. as well as fecal coliforms were evaluated. Overall, Giardia and Vibrio were the most frequently detected potential pathogens, followed by Cryptosporidium and norovirus, while Salmonella was not detected. Sample month, matrix type, and recent precipitation were found to be significant environmental factors for protozoa or host-associated Bacteroidales marker detection, whereas location and water temperature were not. The relative contribution of host-associated markers to total fecal marker concentration was estimated using a Monte Carlo method, with the greatest relative contribution to the Bacteroidales marker concentration coming from human sources, while the remainder of the universal fecal host source signal was uncharacterized by available host-associated assays, consistent with wildlife fecal sources. These findings show how fecal indicator and pathogen monitoring, along with identifying contributing host sources, can provide evidence of coastal pathogen pollution and guidance as to whether to target human and/or animal sources for management.


Assuntos
Baías/microbiologia , Fezes/microbiologia , Sedimentos Geológicos/microbiologia , Microbiologia da Água , Alaska , Animais , Bacteroidetes/isolamento & purificação , Baías/virologia , Cryptosporidium/isolamento & purificação , Ecossistema , Fezes/virologia , Sedimentos Geológicos/virologia , Giardia/isolamento & purificação , Humanos , Método de Monte Carlo , Norovirus/isolamento & purificação , Vibrio/isolamento & purificação , Água , Purificação da Água , Qualidade da Água
20.
J Wildl Dis ; 48(3): 654-68, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22740531

RESUMO

Marine mammals are at risk for infection by fecal-associated zoonotic pathogens when they swim and feed in polluted nearshore marine waters. Because of their tendency to consume 25-30% of their body weight per day in coastal filter-feeding invertebrates, southern sea otters (Enhydra lutris nereis) can act as sentinels of marine ecosystem health in California. Feces from domestic and wildlife species were tested to determine prevalence, potential virulence, and diversity of selected opportunistic enteric bacterial pathogens in the Monterey Bay region. We hypothesized that if sea otters are sentinels of coastal health, and fecal pollution flows from land to sea, then sea otters and terrestrial animals might share the same enteric bacterial species and strains. Twenty-eight percent of fecal samples tested during 2007-2010 were positive for one or more potential pathogens. Campylobacter spp. were isolated most frequently, with an overall prevalence of 11%, followed by Vibrio cholerae (9%), Salmonella spp. (6%), V. parahaemolyticus (5%), and V. alginolyticus (3%). Sea otters were found positive for all target bacteria, exhibiting similar prevalences for Campylobacter and Salmonella spp. but greater prevalences for Vibrio spp. when compared to terrestrial animals. Fifteen Salmonella serotypes were detected, 11 of which were isolated from opossums. This is the first report of sea otter infection by S. enterica Heidelberg, a serotype also associated with human clinical disease. Similar strains of S. enterica Typhimurium were identified in otters, opossums, and gulls, suggesting the possibility of land-sea transfer of enteric bacterial pathogens from terrestrial sources to sea otters.


Assuntos
Infecções por Campylobacter/veterinária , Lontras/microbiologia , Salmonelose Animal/epidemiologia , Vigilância de Evento Sentinela/veterinária , Vibrioses/veterinária , Animais , Animais Domésticos/microbiologia , Animais Selvagens/microbiologia , California/epidemiologia , Campylobacter/isolamento & purificação , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/transmissão , Microbiologia Ambiental , Fezes/microbiologia , Feminino , Humanos , Masculino , Salmonella/isolamento & purificação , Salmonelose Animal/transmissão , Vibrio/isolamento & purificação , Vibrioses/epidemiologia , Vibrioses/transmissão , Microbiologia da Água , Zoonoses
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