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Fig. 1 in Prevalence and geographic distribution of Babesia conradae and detection of Babesia vogeli in free-ranging California coyotes (Canis latrans)

Fig. 1. Base pair differences in a 70 base pair region of the 18S gene from Babesia conradae DNA sequences isolated from California coyotes (Canis latrans) splenic samples collected between 2015 and 2019 compared to published sequence available in GenBank.

Fig. 2 in First detection and low prevalence of Pearsonema spp. in wild raccoons (Procyon lotor) from Central Europe

Fig. 2. Egg of Pearsonema spp. in urine sediment. The egg shows the characteristic features (barrel-shaped with bipolar plugs). Based on their morphology, the eggs were likely to be P. plica (see Discussion). Scale: 20 μm.

Fig. 2. The E in First detection of Echinococcus multilocularis in Bosnia and Herzegovina

Fig. 2. The E. multilocularis adult parasite was determined in the small intestines of the infested red fox using the intestinal scraping technique (stereomicroscopic magnification x100). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 1 in First detection and low prevalence of Pearsonema spp. in wild raccoons (Procyon lotor) from Central Europe

Fig. 1. Sampling areas, sample size (n) and prevalence of Pearonema spp. in wild raccoons (Procyon lotor) from Luxembourg, Poland and Germany. Luxembourg: LUX Luxembourg; Poland: ZL administrative district of Zgorzelecki; Germany: NH Northern Harzvorland, Saxony-Anhalt and administrative district of BZ Bautzen, Saxony; EIC Eichsfeld, Thuringia; PR Prignitz, Brandenburg; GO¨G¨oppingen, Baden-Württemberg.

Fig. 2 in Novel genotypes of Cryptosporidium and Enterocytozoon bieneusi detected in plateau zokors (Myospalax baileyi) from the Tibetan Plateau

Fig. 2. Phylogenetic relationships of E. bieneusi genotypes identified in the present study and other known genotypes deposited on GenBank was inferred by a maximum-likelihood phylogenetic analysis of ITS sequences using the Tamura 3-parameter model with 500 replicates. The Enterocytozoon hepatopenaei (GenBank: KR021167.1) was used as the outgroup. The blue triangle and squares indicate the novel genotypes identified in this study, respectively. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 1 in Novel genotypes of Cryptosporidium and Enterocytozoon bieneusi detected in plateau zokors (Myospalax baileyi) from the Tibetan Plateau

Fig. 1. Phylogenetic relationships of Cryptosporidium sp. genotypes identified in the present study and other known genotypes and species on GenBank was inferred by a maximum-likelihood phylogenetic analysis of SSU rRNA gene sequences using the Tamura 3-parameter model and with 500 replicates. The Eimeria (GenBank: U40264.1) were used as the outgroup. The red circles and squares indicate the novel genotypes identified in this study. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 6 in Plasmodium relictum MSP-1 capture antigen-based ELISA for detection of avian malaria antibodies in African penguins (Spheniscus demersus)

Fig. 6. Analysis of the sensitivity and specificity of the P. relictum MSP-1 capture antigen-based ELISA. Serial dilutions of normal chicken serum and three sera samples from P. relictum-infected penguins (8776, 8783 and 8784) were used for coating the assay wells. Each dilution was performed in triplicate, and the data shown represent means of three independent experiments with standard error bars and levels of statistical significance (****: P <0.0001).

Fig. 4 in Plasmodium relictum MSP-1 capture antigen-based ELISA for detection of avian malaria antibodies in African penguins (Spheniscus demersus)

Fig. 4. Validation of ELISA. The ELISA developed using P. relictum MSP-1 protein was tested with known P. relictum positive penguin serum collected from penguin # 8790 at week 26. Black column represents positive penguin serum. Hatched and white columns represent negative controls containing normal chicken serum and PBS, respectively. Grey column represents the positive reaction containing streptavidin alkaline-phosphatase and its substrate pnitrophenyl phosphate. Each reaction was performed in triplicate, and the data shown represent means of three independent experiments with standard error bars and levels of statistical significance (****: P <0.0001).

Fig. 2 in Plasmodium relictum MSP-1 capture antigen-based ELISA for detection of avian malaria antibodies in African penguins (Spheniscus demersus)

Fig. 2. Verification of the biotinylation of recombinant P. relictum MSP-1. Purified recombinant P. relictum MSP-1 protein was labeled with biotin. Biotinylation of MSP-1 was confirmed using an enzyme (streptavidin alkaline phosphatase)-linking assay and absorbance read at 405 nm indicated the presence of biotinylated MSP-1. Black column represents biotinylated MSP-1 protein. Hatched column and white column represent negative controls containing non-biotinylated MSP-1 and PBS, respectively. Grey column represents the positive control containing streptavidin alkaline-phosphatase (SAP) and its substrate p-nitrophenyl phosphate (PNPP). Each reaction was performed in triplicate, and the data shown represent means of three independent experiments with standard error bars and levels of statistical significance (****: P <0.0001).

Fig. 3 in Plasmodium relictum MSP-1 capture antigen-based ELISA for detection of avian malaria antibodies in African penguins (Spheniscus demersus)

Fig. 3. Biotin-labeled MSP-1 protein titration curve. Serially diluted (300 ng/ μL to 0.003 ng/μL) biotinylated MSP-1 was used for coating the surfaces of the reaction wells overnight at 4 ◦C. The amount of biotinylated MSP-1 immobilized on the surface of the well was proportional to the intensity of the colored product generated which in turn was proportional to the absorbance value measured at 405 nm wavelength. Reactions were performed in triplicate, and the data shown represent means of three independent experiments with standard error bars.

Fig. 7. MSP-1 in Plasmodium relictum MSP-1 capture antigen-based ELISA for detection of avian malaria antibodies in African penguins (Spheniscus demersus)

Fig. 7. MSP-1 capture antigen-based ELISA analysis of 370 sera samples collected from 11 penguins during three consecutive seasons of Spring (March to May), Summer (June to August) and Fall (September to November). (A) ELISA absorbance readings for the 370 sera samples collected from Spring to Fall are arranged in ascending order. Time of collection is indicated by color. Light grey represents Spring, dark grey represents Summer, and black represents Fall. Black dashed line indicates the single cut-off point (0.488) determined by change-point analysis. (B) Left: percentage of positive (A405 ≥ 0.488) and negative (A405 <0.488) sera samples; Right: distribution of positive samples in Spring (March to May), Summer (June to August), and Fall (September to November). (C) Percentage of penguin sera samples that tested positive (A405> = 0.488) in each month from March to November.

Fig. 5. P. relictum MSP-1 in Plasmodium relictum MSP-1 capture antigen-based ELISA for detection of avian malaria antibodies in African penguins (Spheniscus demersus)

Fig. 5. P. relictum MSP-1 capture antigen-based ELISA analysis of test sera from penguins. Sera samples (370 total) from eleven penguins collected from Spring through Fall season were used as test samples in the assay to determine anti-P. relictum antibodies level. Light grey, dark grey, and black columns represent penguin sera samples collected in Spring, Summer, and Fall, respectively. Panel A–K represents ELISA results for individual penguins' sera collected at different time points from Spring to Fall. Panel L represents average ELISA absorbances for all 11 penguins at different sampling points. Each sample was assayed in triplicate, and the data shown represent means of three independent experiments with standard error bars.

Fig. 2 in First detection and molecular identification of Babesia gibsoni and Hepatozoon canis in an Asiatic wild dog (Cuon alpinus) from Thailand

Fig. 2. Neighbor-joining (NJ) tree of the Hepatozoon partial 18S ribosomal RNA (18S rRNA) gene sequence. Hepatozoon canis (MK144332) was amplified from an Asiatic wild dog in Thailand and analyzed for comparison with other Hepatozoon spp. from the GenBank database. The numbers on branches indicate percent bootstrap support based on 1000 bootstrap replications and only bootstrap values ≥ 50% are shown.

Fig. 1 in First detection and molecular identification of Babesia gibsoni and Hepatozoon canis in an Asiatic wild dog (Cuon alpinus) from Thailand

Fig. 1. Neighbor-joining (NJ) tree of the Babesia partial 18S ribosomal RNA (18S rRNA) gene sequence. Babesia gibsoni (MK144331) was amplified from an Asiatic wild dog in Thailand and analyzed for comparison with other Babesia spp. from the GenBank database. The numbers on branches indicate percent bootstrap support based on 1000 bootstrap replications and only bootstrap values ≥ 50% are shown.

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Fig. 1 in Detection of Dirofilaria immitis via integrated serological and molecular analyses in coyotes from Texas, United States

Fig. 1. Texas locations where samples were collected, including Webb, Maverick, Jim Hogg, Haskell, Throckmorton, Hutchinson, Roberts, Reeves, and Loving counties.

Fig. 3 in Sarcocystis calchasi and other Sarcocystidae detected in predatory birds in California, USA

Fig. 3. Phylogenetic tree based on 28S rRNA sequences of Sarcocystis spp. obtained from raptor intestinal samples and reference sequences available in GenBank. The phylogenetic relationships were determined by the Maximum Likelihood method using MEGA-X version 10.2.6 (Kumar et al., 2018). Numbers above or below nodes represent bootstrap confidence values from 500 replicates. The distances were computed using the Tamura-Nei model (Tamura and Nei, 1993). Branch lengths are proportional to sequence divergence and relate to the scale bar (bottom left). GenBank reference sequences are labeled by their accession number and associated information. Raptor sequences are labeled by animal identification number, region amplified (28S), Sarcocystis spp. with the highest homology found by BLASTn, 4-letter alpha-code for the raptor species common name, and NCBI accession number. COHA: Cooper's hawk (Accipiter cooperii), NOGO: northern goshawk (A. gentilis), RSHA: red-shouldered hawk (Buteo lineatus), RTHA: red-tailed hawk (B. jamaicensis), SSHA: sharp-shinned hawk (A. striatus). Note the Eumonospora henryae-like sequence was not included in the analysis. Detailed information for each bird and sequence is presented in Supplementary Table 1.

Fig. 1 in Sarcocystis calchasi and other Sarcocystidae detected in predatory birds in California, USA

Fig. 1. Number of avian carcasses evaluated for Sarcocystis spp. by county in California, USA between 2016 and 2020. Species include American crow (Corvus brachyrhynchos; n = 11), Cooper's hawk (Accipiter cooperii; n = 16), great horned owl (Bubo virginianus; n = 12), northern goshawk (A. gentilis; n = 2), peregrine falcon (Falco peregrinus; n = 3), red-shouldered hawk (Buteo lineatus; n = 12), red-tailed hawk (B. jamaicensis; n = 36), and sharp-shinned hawk (A. striatus; n = 4).

Fig. 2 in Sarcocystis calchasi and other Sarcocystidae detected in predatory birds in California, USA

Fig. 2. Examples of two 28S rRNA chromatograms showing double, overlapping peaks. The chromatogram for northern goshawk (Accipiter gentilis) Z19-0970 (top) from Lassen County, California, USA shows the predominate nucleotide for Sarcocystis columbae (99.7% homology) and secondary nucleotide for S. turdusi (98.7% homology); clear double peaks are patent at positions 219, 234, 241 and 260. The chromatogram for Cooper's hawk (A. cooperii) Z20-0257 (bottom) from Los Angeles County, California, USA shows the predominate nucleotide for S. columbae (99.7% homology) and the secondary nucleotide for S. halieti (100% homology); clear double peaks are patent at positions 249 and 261. Note the marginal presence of noise in the remainder of the sequences.

Fig. 4 in Sarcocystis calchasi and other Sarcocystidae detected in predatory birds in California, USA

Fig. 4. Photomicrograph of intestine from Sarcocystis calchasi-positive Cooper's hawk (Accipiter cooperii) Z17-0062 collected from Los Angeles County, California, USA, showing sporocysts/oocysts (*) in the mucosa. Hematoxylin and eosin stain; scale bar 20 μm.