Nationwide USA re-analysis of amplicon metabarcoding targeting ß-tubulin isoform-1 reveals absence of benzimidazole resistant SNPs in Ancylostoma braziliense, Ancylostoma tubaeforme and Uncinaria stenocephala.

Nationwide sampling by Venkatesan and colleagues (2023) described the resistance status of the canine hookworm, Ancylostoma caninum, to benzimidazoles across the USA via ß-tubulin isotype-1 amplicon metabarcoding. In this study, we aimed to use the existing public amplicon metabarcoding data and mine it for the presence of ß-tubulin isotype-1 sequences that belong to hookworm species other than A. caninum. Through bioinformatics analysis we assigned species to A. caninum, Ancylostoma braziliense, Ancylostoma tubaeforme and Uncinaria stenocephala. All non-A. caninum sequences contained only the benzimidazole susceptible residues of ß-tubulin isotype-1. Using two ß-tubulin isotype-1 metabarcoding sequence data (assay targeting 134 and 167 codons, and assay targeting 198 and 200 codons), 2.0% (6/307) and 2.9% (9/310) individual samples had hookworms other than A. caninum (A. braziliense n = 5, A. tubaeforme n = 4 and U. stenocephala n = 2), respectively. We identified one sample containing A. braziliense in each of the Northeastern region and Midwestern region, and in three samples from the Southern region. Presence of A. tubaeforme in dog faeces is considered as pseudoparasitism. There were no statistically significant regional differences for the distribution of each species, for either of the two assays independently or combined (χ2 tests, P > 0.05). Our work demonstrates the utility of the amplicon metabarcoding for the identification of species through antemortem assays, thus resolving the dilemma of assigning hookworm species based on either post-mortem or egg sizes for the identification of hookworms.

Fatal neural angiostrongyliasis in the Bolivian squirrel monkey (Saimiri boliviensis boliviensis) leading to defining Angiostrongylus cantonensis risk map at a zoo in Australia.

Neural angiostrongyliasis (NA) is a parasitic disease caused by Angiostrongylus cantonensis (rat lungworm). This study presents a case of NA in a captive Bolivian squirrel monkey from a zoo in western Sydney, Australia. The objective was to identify the A. cantonensis cox1 haplotype responsible for the infection and compare its mitochondrial DNA (mtDNA) to known Australian mtDNA. An epidemiological investigation was conducted to assess the risk of infection, focusing on the resident rat population in the zoo. Methods involved trapping rats and collecting rat faeces for Angiostrongylus detection, speciation, and cox1 haplotype confirmation. Various techniques were employed, including necropsy, morphological examination, and molecular methods such as ITS-2 qPCR, cox1 sequencing, and ITS-2 metabarcoding. Cluster analysis of rat faeces distribution and Angiostrongylus detection utilised an equal sampling effort (ESE) approach. Gastropods were collected throughout the study for Angiostrongylus surveillance using a hypersensitive qPCR assay. Results revealed significant clustering of rat faeces near exhibits with fresh food provision and absence of predators. Angiostrongylus-positive faeces were uniformly distributed across the zoo property. Mitochondrial DNA analysis confirmed the presence of the Ac13 haplotype of A. cantonensis in the monkey. Morphology, ITS-2 metabarcoding and partial cox1 sequencing detected only A. cantonensis, with the Ac13 cox1 haplotype predominating. A high prevalence of infection (64%, 9/14) was found in brown rats, with quantification of larvae indicating high shedding rates. Co-infections with both Ac13 and local SYD.1 A. cantonensis cox1 haplotypes were observed. Only three gastropods (all of which were Angiostrongylus-negative) were found in the survey. To minimise the risk of exposure for susceptible species, targeted rodent control was implemented in areas with higher exposure risk. A potential strategy (which requires further exploration) to consider for future zoo design was suggested. This study provides insights into the epidemiology and genetic diversity of A. cantonensis in Australia, emphasising the importance of control measures to prevent future outbreaks.

Spatial epidemiology of cystic echinococcosis in livestock from a hyper-endemic region in southern Chile.

Cystic echinococcosis (CE) is a parasitic zoonosis, caused by the larval stage of Echinococcus granulosus. This disease has a worldwide distribution and is considered an important zoonosis in Chile, especially in regions where raising livestock dominates. We aimed to describe the spatial distribution and risk factors for CE in cattle and sheep slaughtered in the Aysén Region, Chile between 2015 and 2016, to inform disease control in this hyper-endemic region. Clustering and hotspot analyses of CE at the farm-level were performed (Moran's global index, local indicator of spatial autocorrelation (LISA), and the scan statistic Poisson model), and we used spatial interpolation to display areas with a higher risk of CE. Mixed effects logistic regression models were fit to assess the association between municipality of origin and age of animals and CE detected at slaughter, with the farm of origin included as a random effect. Overall, data from 1532 cattle farms (1078 geocoded) and 30,805 cattle were analyzed, with a 64 % and 39 % CE prevalence at the farm and animal-level, respectively. For sheep farms, data from 381 (296 geocoded) farms and 58,223 sheep were analyzed, with a 71 % and 18 % CE prevalence at the farm and animal-level, respectively. Globally, the CE prevalence in both cattle and sheep farms was not clustered. However, we found 14 cattle and 23 sheep farms with high prevalence that were locally clustered. We also detected 6 and 11 clusters in cattle and sheep farms, respectively. Risk of reported CE varied according to municipalities of origin, and the age of animals. This study confirms that the Aysén region is a hyper-endemic region for CE. It demonstrates the usefulness of the traceability of animals slaughtered for understanding the epidemiology of CE. The results suggest that in this hyper-endemic region in which there are limited financial resources available, that intervention activities - at the beginning of a control program - should be focused on farms with high prevalence and identified disease clusters.

Bloodmeal, Host Selection, and Genetic Admixture Analyses of Culex pipiens Complex (Diptera: Culicidae) Mosquitoes in Chicago, IL.

The area in and around Chicago, IL, is a hotspot of West Nile virus activity. The discovery of a Culex pipiens form molestus Forskӓl population in Chicago in 2009 added to speculation that offspring from hybridization between Cx. pipiens f. pipiens L. and f. molestus could show a preference for feeding on humans. We collected blood-fed female mosquitoes (N = 1,023) from eight residential sites and one public park site in Chicago in July and August 2012. Bloodmeal analysis using the COI (cytochrome c oxidase subunit I) gene was performed to ascertain host choice. Almost all (99%) bloodmeals came from birds, with American Robins (Turdus migratorius L.) and House Sparrows (Passer domesticus L.) making up the largest percentage (74% combined). A forage ratio analysis comparing bird species fed upon and available bird species based on point count surveys indicated Northern Cardinals (Cardinalis cardinalis) and American Robins (Turdus migratorius) appeared to be over-utilized, whereas several species were under-utilized. Two human bloodmeals came from Culex pipiens complex mosquitoes. Admixture and population genetic analyses were conducted with 15 microsatellite loci on head and thorax DNA from the collected blood-fed mosquitoes. A modest amount of hybridization was detected between Cx. pipiens f. pipiens and f. molestus, as well as between f. pipiens and Cx. quinquefasciatus Say. Several pure Cx. quinquefasciatus individuals were noted at the two Trumbull Park sites. Our data suggest that Cx. pipiens complex mosquitoes in the Chicago area are not highly introgressed with f. molestus and appear to utilize avian hosts.

Estimating apparent survival of songbirds crossing the Gulf of Mexico during autumn migration.

Many migratory bird species are declining, and the migratory period may limit populations because of the risk in traversing large geographical features during passage. Using automated radio-telemetry, we tracked 139 Swainson's thrushes (Catharus ustulatus) departing coastal Alabama, USA and crossing the Gulf of Mexico to arrive in the Yucatan Peninsula, Mexico during autumn. We estimated apparent survival and examined how extrinsic (weather variables and day of year) and intrinsic (fat load, sex and age) factors influenced survival using a mark-recapture approach. We also examined how favourability of winds for crossing the Gulf varied over the past 25 years. Fat load, day of year and wind profit were important factors in predicting which individuals survived crossing the Gulf. Survival estimates varied with wind profit and fat, but generally, fat birds departing on days with favourable wind profits had an apparent survival probability of greater than 0.90, while lean individuals with no or negative wind profits had less than 0.33. The proportion of favourable nights varied within and among years, but has increased over the last 25 years. While conservation strategies cannot improve extrinsic factors, they can provide opportunities for birds to refuel before crossing large geographical features through protecting and creating high-quality stopover sites.

Swainson's Thrushes do not show strong wind selectivity prior to crossing the Gulf of Mexico.

During long-distance fall migrations, nocturnally migrating Swainson's Thrushes often stop on the northern Gulf of Mexico coast before flying across the Gulf. To minimize energetic costs, trans-Gulf migrants should stop over when they encounter crosswinds or headwinds, and depart with supportive tailwinds. However, time constrained migrants should be less selective, balancing costs of headwinds with benefits of continuing their migrations. To test the hypotheses that birds select supportive winds and that selectivity is mediated by seasonal time constraints, we examined whether local winds affected Swainson's Thrushes' arrival and departure at Ft. Morgan, Alabama, USA at annual, seasonal, and nightly time scales. Additionally, migrants could benefit from forecasting future wind conditions, crossing on nights when winds are consistently supportive across the Gulf, thereby avoiding the potentially lethal consequences of depleting their energetic reserves over water. To test whether birds forecast, we developed a movement model, calculated to what extent departure winds were predictive of future Gulf winds, and tested whether birds responded to predictability. Swainson's Thrushes were only slightly selective and did not appear to forecast. By following the simple rule of avoiding only the strongest headwinds at departure, Swainson's Thrushes could survive the 1500 km flight between Alabama and Veracruz, Mexico.

Fat, weather, and date affect migratory songbirds' departure decisions, routes, and time it takes to cross the Gulf of Mexico.

Approximately two thirds of migratory songbirds in eastern North America negotiate the Gulf of Mexico (GOM), where inclement weather coupled with no refueling or resting opportunities can be lethal. However, decisions made when navigating such features and their consequences remain largely unknown due to technological limitations of tracking small animals over large areas. We used automated radio telemetry to track three songbird species (Red-eyed Vireo, Swainson's Thrush, Wood Thrush) from coastal Alabama to the northern Yucatan Peninsula (YP) during fall migration. Detecting songbirds after crossing ∼1,000 km of open water allowed us to examine intrinsic (age, wing length, fat) and extrinsic (weather, date) variables shaping departure decisions, arrival at the YP, and crossing times. Large fat reserves and low humidity, indicative of beneficial synoptic weather patterns, favored southward departure across the Gulf. Individuals detected in the YP departed with large fat reserves and later in the fall with profitable winds, and flight durations (mean = 22.4 h) were positively related to wind profit. Age was not related to departure behavior, arrival, or travel time. However, vireos negotiated the GOM differently than thrushes, including different departure decisions, lower probability of detection in the YP, and longer crossing times. Defense of winter territories by thrushes but not vireos and species-specific foraging habits may explain the divergent migratory behaviors. Fat reserves appear extremely important to departure decisions and arrival in the YP. As habitat along the GOM is degraded, birds may be limited in their ability to acquire fat to cross the Gulf.

Control of a foot-and-mouth disease epidemic in Argentina.

A major epidemic of foot-and-mouth disease affected Argentina during 2001. The epidemic was controlled by mass-vaccination of the national herd and movement restrictions. The median herd disease reproduction ratio (RH) decreased significantly from 2.4 (before the epidemic was officially recognized) to 1.2 during the mass-vaccination campaign and < 1 following the mass-vaccination campaign. The largest distance between two outbreaks was similar during (1905 km) and after (1890 km) the mass-vaccination. However, after mass-vaccination was completed, the proportion of herd outbreaks clustered decreased from 70.4% to 66.8%, respectively. Although a combination of vaccination and livestock-movement restrictions was effective in controlling the epidemic, 112 herd outbreaks occurred up to 6 months after the end of the mass-vaccination campaign. Mass-vaccination and movement restrictions might be an effective strategy to control FMD; however, the time taken to end large, national epidemics might be > 1 year.

Herd demographics correlated with the spatial distribution of a foot-and-mouth disease epidemic in Buenos Aires province, Argentina.

During a recent foot-and-mouth disease epidemic in Argentina, cattle herds affected in 2001 were located mainly (69%) in Buenos Aires province. The densities of outbreaks (no. of outbreaks per km2) and cattle-demographic variables in the province were estimated using a geographical information system and kernel function. Before the epidemic officially was recognized, the density of outbreaks was correlated (rsp = 0.28-0.47) with the geographic distribution of small (< or =100 cattle), dairy and fattening herds. During the mass-vaccination campaign to control the epidemic (April-July), the density of outbreaks was most strongly correlated (rsp = 0.20-0.25) with the distribution of large (>500 cattle) and breeding herds. After the end of the mass-vaccination campaign, large herds and number of cows were most strongly correlated (rsp = 0.16-0.26) with outbreak density. These relationships might indicate that: (1) the disease spread more rapidly or was more easily detected in intensive production systems at the beginning of the epidemic; (2) vaccination and other control methods applied were less effective in large, semi-intensive production systems; (3) incomplete vaccine protection was responsible for herd outbreaks that occurred after the end of the mass-vaccination campaign.

Use of spatial statistics and monitoring data to identify clustering of bovine tuberculosis in Argentina.

The spatial distribution of endemic bovine tuberculosis (TB) in Argentine cattle herds was described using recorded information on the detection of TB-like lesions in cattle slaughtered between March 1995 and 1997 at 126 slaughterhouses with federal inspection. Approximately, 47% (9472396 cattle) of the estimated total number of cattle slaughtered in Argentina during this period was included in the study. Information on the number of cattle per source herd consigned to slaughter, number of cattle with TB-like lesions per herd and the geographical location of counties from which cattle originated was used to investigate spatial clustering of TB. Overall, no evidence of clustering of TB prevalence by county was detected (Moran's autocorrelation statistic I=0.009, P=0.089). However, first- (Cuzick and Edwards' test statistic, T(k)=87, P=0.036) and second-order (T(k)=170, P=0.038) nearest-neighbor case-counties (TB prevalence>median prevalence of all counties, 6.7%) were clustered. Using the spatial scan test based on a Bernoulli model, the most-likely cluster (P=0.001) identified during the study period included 5793 cases of TB (5.2 per 1000 km(2)) in five counties. This cluster coincided with Santa Fe Province, which contains 21% of all dairy cows in Argentina and accounts for 34% of the country's milk production. Several secondary clusters of TB-also located in dairy districts-were identified. Study results demonstrate that bovine TB is clustered in Argentina, and these clusters coincide with dairy cattle production. Identification of clustering can assist efforts to eradicate bovine TB from Argentina. Further spatial investigations need to focus on the reasons why TB is clustered in Argentina. In particular, the relationship between TB clustering and management practices-such as grazing density and production systems-need to be described to assist in the development of disease-control programs. The use of spatial statistics and geographical information systems could meet these needs.

Simulation model of within-herd transmission of bovine tuberculosis in Argentine dairy herds.

Transmission of bovine tuberculosis was quantified in three dairy herds located in south Santa Fe Province, Argentina. Using estimates of Mycobacterium bovis transmission (beta) and a Reed-Frost simulation model, the prevalence of tuberculosis infection in the study herds over time was investigated. The Reed-Frost model was modified by incorporating randomness in both beta and the incubation period (alpha) of M. bovis. The mean estimated herd beta was 2.2 infective contacts per year and did not differ significantly between the study herds. Modeling alpha as Poisson distributed (mean 24 months) best fit the observed prevalences. Infection was predicted by the model either to spread quickly (<10 years) within a herd and reach a high prevalence (>50%), or to persist at a low prevalence (<5-10%). The model was robust, predictions were realistic and the mean beta estimated was consistent with previous studies of bovine tuberculosis.

Simulation-model evaluation of bovine tuberculosis-eradication strategies in Argentine dairy herds.

We used stochastic modification of the Reed-Frost model to assess the impact of 14 different eradication strategies on bovine tuberculosis, under three scenarios of disease introduction, in Argentine dairy herds. All strategies investigated were based on a test-and-cull approach using either the caudal-fold test (CFT), the single cervical test (SCT), the gamma-interferon (gamma-IFN) test or a combination of these tests. The maximum, minimum and most-likely sensitivity and specificity were investigated in three scenarios simulating different epidemiological conditions. Simulation results were highly variable; therefore, it is difficult to predict the effect of disease-control strategies within individual herds. On average, the use of the SCT was less efficient in eradicating tuberculosis from the simulated herd than the CFT. Eradication would be achieved most efficiently by strategies in which the CFT was used assuming maximum possible sensitivity and specificity (difficult to achieve in the field) and/or the gamma-IFN test-which has both economical and logistical limitations to its widespread application in Argentina. When disease-control was simulated in situations in which herd tuberculosis prevalence is > or = 22%, all strategies we simulated were less efficient than herd depopulation. Considering that Argentine dairy producers are not compensated financially for cattle culled because of tuberculosis, eradication strategies currently used in the Argentine national tuberculosis eradication might not succeed.