Isolation and characterization of pathogenic leptospires associated with cattle

2020

Sci Rep

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The causative agent of leptospirosis includes multiple serovars and species of pathogenic leptospires that are excreted via urine from reservoir hosts of infection. Primary isolation takes weeks to months, and is limited to semi-solid media at 28-30 °C. Here we present an alternative media formulation, HAN, compared to commercially available EMJH and the more specialized T80/40/LH media formulations, in semi-solid and liquid compositions, for the primary isolation of two diverse species and serovars of pathogenic leptospires directly from host kidney tissue. All three media types supported the isolation and propagation of L. interrogans serovar Copenhageni strain IC:20:001 in semi-solid media at 29 °C. However, only HAN and T80/40/LH supported the growth of L. borgpetersenii serovar Hardjo strain HB15B203 at 29 °C. In addition, HAN supported primary isolation at 37 °C. Both T80/40/LH and HAN supported primary isolation of strain IC:20:001 in liquid media at 29 °C but only HAN supported growth of strain HB15B203 in liquid media, at both 29 and 37 °C. HAN media supports the primary isolation of fastidious pathogenic leptospires directly from infected host tissue at either 29 or 37 °C: this formulation represents a more defined media for the continued optimization of growth factors required to support the primary isolation of the large and diverse range of species and serovars within the genus Leptospira circulating within domestic and wild animal populations.

mentioning

confidence: 99%

Isolation and propagation of leptospires at 37 °C directly from the mammalian host

Hornsby

Alt

2020

Sci Rep

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“…Original magnification 10,000×. (Natarajaseenivasan et al, 2005;Ramirez-Ramirez et al, 2015) SP (Boey et al, 2019) SP (Campos et al, 2017) A (Ojeda et al, 2018;Loureiro and Lilenbaum, 2020) SP (Hamond et al, 2015;Simbizi et al, 2016) A (Hamond et al, 2015) R (Gay et al, 2014) SP (Lee et al, 2017) A (Mitchell et al, 1966) A (Adin and Cowgill, 2000;Greenlee et al, 2005) SP (Goldstein et al, 2006) A (Srinivas et al, 2013;Techawiwattanaboon et al, 2019) Tarassovi A (Maze et al, 2016) SP (Boey et al, 2019) SP (Campos et al, 2017) SP (Simbizi et al, 2016) SP (Lee et al, 2017) R SP (Barmettler et al, 2011) NE Hardjo A (Bolin and Koellner, 1988;Ezeh et al, 1991) R SP (Boey et al, 2019) R* (Ellis et al, 1981;Ellis et al, 1986;Rajala et al, 2017;Loureiro and Lilenbaum, 2020) SP (Campos et al, 2017;Nally et al, 2018a) A (Hamond et al, 2015) SP (Simbizi et al, 2016) SP (Hathaway et al, 1983;Lee et al, 2017) SP (Goldstein et al, 2006;Harland et al, 2013) A (Zuerner et a...…”

Section: The Complicated Relationship Of Leptospira and Various Hostmentioning

confidence: 99%

“…Antibody production may still be induced in reservoir hosts, but not necessarily. In cattle, there is ample evidence of culture positive animals that were seronegative by MAT (Miller et al, 1991;Nally et al, 2018a), and there are also examples of infected turtles that are PCR positive/MAT (microscopic agglutination test) negative (Oliveira et al, 2016). Bovine neutrophils co-cultured with L. borgpetersenii (serovar Hardjo) also minimally produce Neutrophil Extracellular Traps (NET) compared to Escherichia coli and PMA controls (Wilson-Welder et al, 2016).…”

Section: Model Reservoir Hosts: the Cow And The Ratmentioning

confidence: 99%

Investigating the Immunological and Biological Equilibrium of Reservoir Hosts and Pathogenic Leptospira: Balancing the Solution to an Acute Problem?

Putz

2020

Front. Microbiol.

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Leptospirosis is a devastating zoonotic disease affecting people and animals across the globe. Pathogenic leptospires are excreted in urine of reservoir hosts which directly or indirectly leads to continued disease transmission, via contact with mucous membranes or a breach of the skin barrier of another host. Human fatalities approach 60,000 deaths per annum; though most vertebrates are susceptible to leptospirosis, complex interactions between host species and serovars of Leptospira can yield disease phenotypes that vary from asymptomatic shedding in reservoir hosts, to multiorgan failure in incidental hosts. Clinical symptoms of acute leptospirosis reflect the diverse range of pathogenic species and serovars that cause infection, the level of exposure, and the relationship of the pathogen with the given host. However, in all cases, pathogenic Leptospira are excreted into the environment via urine from reservoir hosts which are uniformly recognized as asymptomatic carriers. Therefore, the reservoir host serves as the cornerstone of persistent disease transmission. Although bacterin vaccines can be used to abate renal carriage and excretion in domestic animal species, there is an urgent need to advance our understanding of immune-mediated host-pathogen interactions that facilitate persistent asymptomatic carriage. This review summarizes the current understanding of host-pathogen interactions in the reservoir host and prioritizes research to unravel mechanisms that allow for colonization but not destruction of the host. This information is required to understand, and ultimately control, the transmission of pathogenic Leptospira.

“…Bovine leptospirosis can result in abortion, stillbirth, premature birth, reproductive failure and milk drop syndrome [ 4 ]. Cattle are susceptible to infection with multiple Leptospira species and serovars including L. borgpetersenii serovar Hardjo, L. interrogans serovar Pomona, L. kirschneri serovar Grippotyphosa and L. noguchii [ 10 , 11 , 12 ]. However, the most prominent serovar associated with cattle is Hardjo, which causes reproductive failure [ 8 , 11 , 13 ].…”

Section: Introductionmentioning

confidence: 99%

“…Seroprevalence studies indicate that up to 49% of cattle are exposed to pathogenic serovars [ 11 ]. Seronegative animals may also excrete Leptospira [ 11 , 12 ].…”

Section: Introductionmentioning

confidence: 99%

Comparison of Real-Time PCR, Bacteriologic Culture and Fluorescent Antibody Test for the Detection of Leptospira borgpetersenii in Urine of Naturally Infected Cattle

Ahmed

Putz

et al. 2020

Veterinary Sciences

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Cattle are susceptible to infection with multiple serovars of pathogenic leptospires, resulting in abortion, stillbirth, premature birth, reproductive failure and milk drop syndrome. Cattle also act as a reservoir host for L. borgpetersenii serovar Hardjo which is excreted from renal tubules via urine into the environment where it persists in suitable moist conditions. Our previous work demonstrated that 7% of urine samples from beef cattle were positive for L. borgpetersenii serovar Hardjo by culture and/or the fluorescent antibody test (FAT). In this study, a real-time PCR (rtPCR) assay was applied to determine the relative performance of rtPCR based detection of L. borgpetersenii serovar Hardjo compared to previously reported culture and FAT techniques. Of 42 bovine urine samples positive for leptospires by culture and/or FAT, 60% (25/42) were positive by rtPCR. Of 22 culture-positive samples, 91% (20/22) were rtPCR-positive. Of 32 FAT-positive samples, 50% (16/32) were rtPCR-positive. For 10 samples that were culture-positive but FAT-negative, 90% (9/10) were rtPCR-positive. For 20 samples that were FAT-positive but culture-negative, 25% (5/20) were rtPCR-positive. Collectively, these results indicate that no single assay is optimal, and the use of more than one assay to detect leptospires in urine from naturally infected cattle is recommended.