Babesia equi Field Isolates Cultured from Horse Blood Using a Microcentrifuge Method

Holman, Patricia J.; Becú, Teótimo; Bakos, E; Polledo, Gonzalo; Cruz, D; Wagner, G. G.

doi:10.2307/3284572

Cited by 16 publications

(16 citation statements)

References 3 publications

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“…In their study on the use of in vitro culture methods to isolate B. equi, Holman et al were unable to subculture 30% of the isolated parasites [14]. However, several results indicate that such a strong selection did not occur in our conditions.…”

mentioning

confidence: 69%

“…An in vitro isolation procedure of Babesia from carrier animals has been reported in the case of B. equi [14,22] and B. caballi [13] from horses and Babesia sp. from free-ranging artiodactylids [19].…”

mentioning

confidence: 99%

“…In one of these studies, 2.5 µL of packed erythrocytes were sufficient to initiate the B. equi culture, demonstrating the good sensitivity of the method [22]. However, the selective effect of the in vitro culture method on the parasite is sometimes very high [14].…”

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confidence: 99%

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Isolation of Babesia divergens from carrier cattle blood using in vitro culture

2004

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-Babesia divergens, the main causative agent of bovine babesiosis in Western Europe, was isolated from naturally infected cattle. Ninety-six blood samples were examined by means of an in vitro culture technique in sheep erythrocytes: 19 of them were collected from animals in the acute phase of the disease with visible parasitemia on blood smears, while the 77 remaining animals showed no microscopically detectable parasites. B. divergens was cultured from the 19 first blood samples as well as from 31 samples collected from asymptomatic animals. The time period before parasites could be detected in the culture varied in the latter samples from 6 to 20 days. The effects of sampling condition (anticoagulant used) and storage length were tested. A good correlation was obtained between immunofluorescent antibody test and culture, with identical results (positive or negative) for 89.6% of the samples collected from asymptomatic animals. The sensitivity of the in vitro culture method was determined and was about 10 parasites/mL of whole blood from three independent experiments performed with three different isolates, confirming its suitability to detect and culture diverse B. divergens isolates from carrier cattle. The parasites could indeed be isolated 9 months after the acute babesiosis phase in the blood of naturally infected animals. The 50 isolates collected in this study were successfully subcultured, cryopreserved and resuscitated using the same culture medium. The in vitro isolation of B. divergens from asymptomatic carrier cattle was achieved and will allow the analysis of parasite diversity within cattle herds. Babesia divergens / carrier cattle / in vitro culture / isolation

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confidence: 69%

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confidence: 99%

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Isolation of Babesia divergens from carrier cattle blood using in vitro culture

2004

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“…Bose et al (1995) reported the low sensitivity of this technique and suggest its use only in acute cases of the disease. Holman et al (1998) suggested that negative results in stained blood smears could be due to time delay and handling between blood collection and slide preparation. Therefore, it should be mentioned therefore, that in the present study, the blood was immediately processed after the return from the treadmill, implicating in intervals of approximately three hours, a fact that would not justify the occurrence of false-negative results.…”

Section: Discussionmentioning

confidence: 99%

“…Nested PCR has also been proven to be more sensitive in the detection of T. equi parasites than traditional microscope procedures, as it is able to detect the parasite in blood with an equivalent 0.000006% parasitemia (Nicolaiewsky et al, 2001). Additionally, it has been shown that in vitro techniques are capable of successfully detecting the carrier status of horses suspected of harboring T. equi (Holman et al, 1993;Zweygarth et al, 1997;Holman et al, 1998). Results from these studies suggests that in vitro culture could contribute to the identification of carrier animals and complement other methods of parasite detection, such as microscopy, serology, or PCR.…”

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confidence: 99%

In vitro culture, PCR , and nested PCR for the detection of Theileria equi in horses submitted to exercise

Baldani

Canola

Neto

et al. 2008

Arq. Bras. Med. Vet. Zootec.

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This study compared the usefulness of in vitro culture, PCR, and nested PCR for the diagnosis of Theileria equi in horses submitted to stress during exercise. Blood samples from 15 apparently healthy horses, previously conditioned to a high-speed equine treadmill, were taken prior to and after exercise. The animals were divided into two experimental groups: 30-day training schedule (G1) and 90-day training schedule (G2). Statistical analysis was performed using a chi-square test and kappa statistic was used in order to assess agreement. No significant difference was observed between samples collected at resting or after exercise. In G1, merozoites of T. equi were detected in the blood smears of four horses before in vitro culture, whereas 14 samples were positive, confirmed by culture. In G2, five and 11 horses were positive before and after culture, respectively. No PCR amplified product was observed in any of the tested animals although the PCR system based on the 16S rRNA gene of T. equi detected DNA in blood with an equivalent 8x10-5% parasitaemia. The nested PCR based on the T. equi merozoite antigen gene (EMA-1) allowed the visualization of amplified products in all the horses. Therefore, nested PCR should be considered as a means of detection of sub-clinical T. equi infections and in vitro culture could be used as a complement to other methods of diagnosis.

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Genotypically unique Babesia spp. isolated from reindeer (Rangifer tarandus tarandus) in the United States

et al. 2002

Parasitol Res

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Two morphologically dissimilar Babesia spp. were cultured from reindeer (Rangifer tarandus tarandus) in Placer County, Calif. The smaller isolate, designated RD61, was morphologically similar to Babesia odocoilei. Serum from RD61-infected reindeer reacted equally strongly to B. odocoilei and RD61 parasites in the indirect fluorescent antibody (IFA) test. Small subunit ribosomal RNA (SSU rRNA) gene-sequence analysis showed 99.0% identity to that of B. odocoilei. The larger piroplasm, designated RD63, resembled larger babesia organisms, such as Babesia caballi and Babesia bigemina. Serum from RD63-infected reindeer also reacted with both B. odocoilei and RD61 parasites in the indirect fluorescent antibody test. The SSU rRNA gene showed 94.2% identity to that of B. bigemina. Further studies are needed to determine whether these parasites are the same as the Babesia spp. previously documented in Siberian reindeer.

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Babesia equi Field Isolates Cultured from Horse Blood Using a Microcentrifuge Method

Cited by 16 publications

References 3 publications

Isolation of Babesia divergens from carrier cattle blood using in vitro culture

Isolation of Babesia divergens from carrier cattle blood using in vitro culture

In vitro culture, PCR , and nested PCR for the detection of Theileria equi in horses submitted to exercise

Genotypically unique Babesia spp. isolated from reindeer (Rangifer tarandus tarandus) in the United States

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