Recent insights into alteration of red blood cells by Babesia bovis: moovin’ forward

Gohil, Sejal; Kats, Lev; Sturm, Angelika; Cooke, Brian M.

doi:10.1016/j.pt.2010.06.012

Cited by 60 publications

(59 citation statements)

References 72 publications

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“…The present report describes the potential application of the fluorescence-based method for antibabesial drug screening using B. bovis, which is thought to be the most virulent Babesia parasite to cause clinical babesiosis among cattle (18). In preliminary experiments, linearity between the fluorescence signals emitted from SG I-treated B. bovis parasites and the microscopy-based parasitemia was observed with high r 2 values.…”

Section: Discussionmentioning

confidence: 83%

Evaluation of a Fluorescence-Based Method for Antibabesial Drug Screening

Guswanto

Sivakumar

Rizk

et al. 2014

Antimicrob Agents Chemother

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In vitro evaluation of chemotherapeutic agents against Babesia and Theileria parasites has become routine, and the effectiveness of these chemicals is usually determined by comparing the parasitemia dynamics of untreated and treated parasites. Although microscopy is widely used to calculate parasitemia, several disadvantages are associated with this technique. The present study evaluated a fluorescence-based method using SYBR green I stain (SG I) to screen antibabesial agents in in vitro cultures of Babesia bovis. The linearity between relative fluorescence units (RFU) and parasitemia was found to be well correlated with a 0.9944 goodness-of-fit (r 2 ) value. Subsequently, 50% inhibitory concentration (IC 50 ) values were calculated for 3 antiprotozoan agents, diminazene aceturate, nimbolide, and gedunin, by this method. For diminazene aceturate and nimbolide, the IC 50 s determined by the fluorescence-based method (408 nM and 8.13 M, respectively) and microscopy (400.3 nM and 9.4 M, respectively) were in agreement. Furthermore, the IC 50 of gedunin determined by the fluorescence-based method (19 M) was similar to the recently described microscopy-based value (21.7 M) for B. bovis. Additionally, the Z= factor (0.80 to 0.90), signal-to-noise (S/N) ratio (44.15 to 87.64), coefficient of variation at the maximum signal (%CVmax) (0.50 to 2.85), and coefficient of variation at the minimum signal (%CVmin) (1.23 to 2.21) calculated for the fluorescence method using diminazene aceturate were comparable to those previously determined in malaria research for this assay. These findings suggest that the fluorescence-based method might be useful for antibabesial drug screening and may have potential to be developed into a high-throughput screening (HTS) assay. Babesiosis, which is caused by Babesia parasites, has a deleterious effect on the health status of cattle (1). Early diagnosis and treatment with effective antiprotozoan agents are essential for the fast recovery of affected animals (2, 3). The development of drug resistance to and toxic side effects of the drugs currently available for treating babesiosis indicate the importance of introducing new alternative treatment options in veterinary practice (4). Potential novel antibabesial agents are usually evaluated in vitro before clinical trials are conducted. At present, the growth-inhibiting effects of the drug candidates are determined based on parasitemia dynamics, which usually involves microscopic examination of Giemsa-stained thin blood smears of treated and untreated parasites (5, 6). However, microscopy that requires good-quality smears is a time-consuming technique and is unsuitable for mass screening of potential drug candidates. In addition, significant differences in parasitemia estimated by different personnel may be found.Incorporation 3 H]isoleucine, has been a common method in in vitro antimalarial drug screening (7,8). This method is slowly being replaced by a fluorescence-based method because the former is costly, requires numerous procedures, and is c...

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Section: Discussionmentioning

confidence: 83%

Evaluation of a Fluorescence-Based Method for Antibabesial Drug Screening

Guswanto

Sivakumar

Rizk

et al. 2014

Antimicrob Agents Chemother

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“…When taking a blood meal, infected ticks infect a mammalian host with sporozoites (94). These sporozoites enter erythrocytes and reproduce through asynchronous binary fission, resulting in two, or sometimes four, merozoites (95,96). Once present in a reservoir host, parasites will develop into male and female gametes (95,96).…”

Section: Babesiosis Life Cycle and Mechanisms Of Virulencementioning

confidence: 99%

“…These sporozoites enter erythrocytes and reproduce through asynchronous binary fission, resulting in two, or sometimes four, merozoites (95,96). Once present in a reservoir host, parasites will develop into male and female gametes (95,96). Zoonotic Babesia species have diverse reservoirs, including the white-footed mouse, cattle, wild ruminants, canids, shrews, and, possibly, cottontail rabbits (Table 1).…”

Section: Babesiosis Life Cycle and Mechanisms Of Virulencementioning

confidence: 99%

“…In addition to VMSA, Babesia spp. produce variable erythrocyte surface antigen (VESA), which is transported to the erythrocyte surface much like Plasmodium knob proteins, causing adhesion to the endothelial cells of small vessels (95). This serves the function of sequestering infected erythrocytes in the peripheral microvasculature and away from the spleen.…”

Section: Unverified Cases Reviewed Previously By Gorenflot Et Al (125)mentioning

confidence: 99%

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Transmission and Epidemiology of Zoonotic Protozoal Diseases of Companion Animals

2013

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SUMMARYOver 77 million dogs and 93 million cats share our households in the United States. Multiple studies have demonstrated the importance of pets in their owners' physical and mental health. Given the large number of companion animals in the United States and the proximity and bond of these animals with their owners, understanding and preventing the diseases that these companions bring with them are of paramount importance. Zoonotic protozoal parasites, including toxoplasmosis, Chagas' disease, babesiosis, giardiasis, and leishmaniasis, can cause insidious infections, with asymptomatic animals being capable of transmitting disease.GiardiaandToxoplasma gondii, endemic to the United States, have high prevalences in companion animals.LeishmaniaandTrypanosoma cruziare found regionally within the United States. These diseases have lower prevalences but are significant sources of human disease globally and are expanding their companion animal distribution. Thankfully, healthy individuals in the United States are protected by intact immune systems and bolstered by good nutrition, sanitation, and hygiene. Immunocompromised individuals, including the growing number of obese and/or diabetic people, are at a much higher risk of developing zoonoses. Awareness of these often neglected diseases in all health communities is important for protecting pets and owners. To provide this awareness, this review is focused on zoonotic protozoal mechanisms of virulence, epidemiology, and the transmission of pathogens of consequence to pet owners in the United States.

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“…They induce structural and/ or antigenic changes in erythrocytes membrane, creating binding sites for receptors expressed in endothelial cells (Gohil et al 2010).…”

Section: Resumo-[babesia Bovis: Expressão De Moléculas De Adesão Em mentioning

confidence: 99%

Babesia bovis: expression of adhesion molecules in bovine umbilical endothelial cells stimulated with plasma from infected cattle

et al. 2014

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Pesq. Vet. Bras. 34(10):937-941, outubro 2014 937 RESUMO.-[Babesia bovis: expressão de moléculas de adesão em células endoteliais de cordão umbilical de bovinos estimuladas com plasma de bovinos infectados.] Dez bezerros machos, da raça Jersey, com 1 ano de idade com baços "in situ", sorológica e parasitologicamente livres de Babesia, foram mantidos em baias individuais no isolamento a prova de artrópodes do Depto de Veterinária desde o nascimento e ao longo de toda a experimentação. Os animais foram divididos aleatoriamente em dois grupos. Cinco animais (grupo A) foram inoculados por via intravenosa com 6,6 x10 7 hemácias parasitados com amostra patogênica de Babesia bovis (BboUFV -1 7 ª passagem) , para a determinação subseqüente "ex vivo" da expressão de moléculas de adesão . Cinco animais não inoculados (Grupo B ) foram utilizados como controlo negativo . A expressão de moléculas de adesão ICAM -1, VCAM , PECAM -1, E -selectina e trombospondina ( TSP ) foi medida em células endoteliais da veia umbilical de bovinos (BUVECs). As células endoteliais estimuladas com um pool de plasma proveniente de animais infectados com BboUFV -1 7 ª passagem tinham uma imunocoloração muito mais intensa de ICAM -1 , VCAM , PECAM -1 de E -selectina e de TSP , em comparação com as células que não receberam o estímulo . Os resultados sugerem que as citocinas pró-inflamatórias liberados na fase aguda da babesiose pode estar envolvida na expressão de moléculas de adesão , implicando , assim, elas na fisiopatologia da babesiose causada por B. bovis. Ten male, 12-month-old Jersey with intact spleens, serologically and parasitologically free from Babesia were housed individually in an arthropod-free isolation system from birth and throughout entire experiment. The animals were randomly divided into two groups. Five animals (group A) were intravenously inoculated with 6.6 X10 7 red blood cells parasitized with pathogenic sample of Babesia bovis (passage 7 BboUFV-1), for the subsequent "ex vivo" determination of the expression of adhesion molecules. Five non-inoculated animals (group B) were used as the negative control. The expression of the adhesion molecules ICAM-1, VCAM, PECAM-1 E-selectin and thrombospondin (TSP) was measured in bovine umbilical vein endothelial cells (BUVECs). The endothelial cells stimulated with a pool of plasma from animals infected with the BboUFV-1 7 th passage sample had a much more intense immunostaining of ICAM-1, VCAM, PECAM-1 E-selectin and TSP, compared to the cells which did not received the stimulus. The results suggest that proinflammatory cytokines released in the acute phase of babesiosis may be involved in the expression of adhesion molecules thereby implicating them in the pathophysiology of babesiosis caused by B. bovis.

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Recent insights into alteration of red blood cells by Babesia bovis: moovin’ forward

Cited by 60 publications

References 72 publications

Evaluation of a Fluorescence-Based Method for Antibabesial Drug Screening

Evaluation of a Fluorescence-Based Method for Antibabesial Drug Screening

Transmission and Epidemiology of Zoonotic Protozoal Diseases of Companion Animals

Babesia bovis: expression of adhesion molecules in bovine umbilical endothelial cells stimulated with plasma from infected cattle

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