Connectivity-based risk ranking of infectious salmon anaemia virus (ISAv) outbreaks for targeted surveillance planning in Canada and the USA.

Infectious salmon anaemia (ISA) can be a serious viral disease of farmed Atlantic salmon (Salmo salar). A tool to rank susceptible farms based on the risk of ISA virus (ISAv) infection spread from infectious farms after initial incursion or re-occurrence in an endemic area, can help guide monitoring and surveillance activities. Such a tool could also support the response strategy to contain virus spread, given available resources. We developed a tool to rank ISAv infection risks using seaway distance and hydrodynamic information separately and combined. The models were validated using 2002-2004 ISAv outbreak data for 30 farms (24 in New Brunswick, Canada and 6 in Maine, United States). Time sequence of infection spread was determined from the outbreak data that included monthly infection status of the cages on these farms. The first infected farm was considered as the index site for potential spread of ISAv to all other farms. To assess the risk of ISAv spreading to susceptible farms, the second and subsequent infected farms were identified using the farm status in the given time period and all infected farms from the previous time periods. Using the three models (hydrodynamic only, seaway-distance, and combined hydrodynamic-seaway-distance based models), we ranked susceptible farms within each time interval by adding the transmission risks from surrounding infected farms and sorting them from highest to lowest. To explore the potential efficiency of targeted sampling, we converted rankings to percentiles and assessed the model's predictive performance by comparing farms identified as high risk based on the rank with those that were infected during the next time interval as observed in the outbreak data. The overall predictive ability of the models was compared using area under the ROC curve (AUC). Farms that become infected in the next period were always within the top 65% of the rank predicted by our models. The overall predictive ability of the combined (hydrodynamic-seaway-distance based model) model (AUC = 0.833) was similar to the model that only used seaway distance (AUC = 0.827). Such models can aid in effective surveillance planning by balancing coverage (number of farms included in surveillance) against the desired level of confidence of including all farms that become infected in the next time period. Our results suggest that 100% of the farms that become infected in the next time period could be targeted in a surveillance program, although at a significant cost of including many false positives.

Evaluating the effect of synchronized sea lice treatments in Chile.

The sea louse is considered an important ectoparasite that affects farmed salmonids around the world. Sea lice control relies heavily on pharmacological treatments in several salmon-producing countries, including Chile. Among options for drug administration, immersion treatments represent the majority of antiparasitic control strategies used in Chile. As a topical procedure, immersion treatments do not induce a long lasting effect; therefore, re-infestation from neighbouring farms may undermine their efficacy. Synchronization of treatments has been proposed as a strategy to improve immersion treatment performance, but it has not been evaluated so far. Using a repeated-measures linear mixed-effect model, we evaluated the impact of treatment synchronization of neighbouring farms (within 10km seaway distance) on the adult lice mean abundance from weeks 2 to 8 post-treatment on rainbow trout and Atlantic salmon farms in Chile, while controlling for external and internal sources of lice before the treatments, and also for environmental and fish-related variables. Results indicate that treatment synchronization was significantly associated with lower adult lice levels from weeks 5 to 7 after treatment. This relationship appeared to be linear, suggesting that higher levels of synchronization may result in lower adult sea lice levels during these weeks. These findings suggest that synchronization can improve the performance of immersion delousing treatments by keeping sea lice levels low for a longer period of time. Our results may be applicable to other regions of the world where immersion treatments are widely used.

Effect of timing of count events on estimates of sea lice abundance and interpretation of effectiveness following bath treatments.

Effectiveness of sea lice bath treatment is often assessed by comparing pre- and post-treatment counts. However, in practice, the post-treatment counting window varies from the day of treatment to several days after treatment. In this study, we assess the effect of post-treatment lag time on sea lice abundance estimates after chemical bath treatment using data from the sea lice data management program (Fish-iTrends) between 2010 and 2014. Data on two life stages, (i) adult female (AF) and (ii) pre-adult and adult male (PAAM), were aggregated at the cage level and log-transformed. Average sea lice counts by post-treatment lag time were computed for AF and PAAM and compared relative to treatment day, using linear mixed models. There were 720 observations (treatment events) that uniquely matched pre- and post-treatment counts from 53 farms. Lag time had a significant effect on the estimated sea lice abundance, which was influenced by season and pre-treatment sea lice levels. During summer, sea lice were at a minimum when counted 1 day post-treatment irrespective of pre-treatment sea lice levels, whereas in the spring and autumn, low levels were observed for PAAM over a longer interval of time, provided the pre-treatment sea lice levels were >5-10.

Evaluating bath treatment effectiveness in the control of sea lice burdens on Atlantic salmon in New Brunswick, Canada.

The use of medicinal bath treatment for sea lice is becoming more common, due to increasing resistance to in-feed treatments with emamectin benzoate. Common treatment modalities in New Brunswick, Canada, include Salmosan administered by tarpaulin or wellboat, and Paramove administered by wellboat. In this study, we assessed the effectiveness of these treatment modalities in the field between 2010 and 2015 using a web-based sea lice data management system (Fish-iTrends© ). Effectiveness was evaluated for adult female (AF) and for pre-adult and adult male (PAAM) life stages separately. We also investigated the impact of variability in pretreatment lead and post-treatment lag time on effectiveness measures. There were 1185 treatment events at 57 farms that uniquely matched our pre- and post-treatment count criteria. The effectiveness of treatment modality was significantly influenced by season, pretreatment level of sea lice and by lead and lag times. In summer, Salmosan administered by tarpaulin had the greatest effectiveness on both AF and PAAM, when pretreatment levels were above 10 sea lice; whereas in autumn, the performance of treatment modalities varied significantly, depending on the pretreatment levels for the life stages. Ignoring the lead or lag time effect generally resulted in an underestimation of treatment effectiveness.

Variation in pre-treatment count lead time and its effect on baseline estimates of cage-level sea lice abundance.

Treatment efficacy studies typically use pre-treatment sea lice abundance as the baseline. However, the pre-treatment counting window often varies from the day of treatment to several days before treatment. We assessed the effect of lead time on baseline estimates, using historical data (2010-14) from a sea lice data management programme (Fish-iTrends). Data were aggregated at the cage level for three life stages: (i) chalimus, (ii) pre-adult and adult male and (iii) adult female. Sea lice counts were log-transformed, and mean counts by lead time relative to treatment day were computed and compared separately for each life stage, using linear mixed models. There were 1,658 observations (treatment events) from 56 sites in 5 Bay Management Areas. Our study showed that lead time had a significant effect on the estimated sea lice abundance, which was moderated by season. During the late summer and autumn periods, counting on the day of treatment gave significantly higher values than other days and would be a more appropriate baseline estimate, while during spring and early summer abundance estimates were comparable among counts within 5 days of treatment. A season-based lead time window may be most appropriate when estimating baseline sea lice levels.

Effects of surgical and chemical sterilization on the behavior of free-roaming male dogs in Puerto Natales, Chile.

Population management of free-roaming domestic dogs (Canis lupus familiaris) is of interest due to the threat these animals pose to people, other animals and the environment. Current sterilization procedures for male dogs include surgical and chemical methods. However, little is known about how these procedures affect their behavior. The primary objective of this study was to investigate changes in selected behaviors following chemical and surgical sterilization in a male free-roaming dog (FRD) population in southern Chile. We also examined the association between serum testosterone levels and behaviors thought to be influenced by circulating androgens. A total of 174 dogs were randomly assigned to either a surgical or chemical sterilization group, or a control group. At the onset of the intervention period, 119 dogs remained and 102 dogs successfully completed the study. Each dog was monitored pre- and post-intervention using video recordings, GPS collars, and blood samples for the measurement of testosterone. Analysis of behavior revealed that surgically castrated dogs showed no reduction of sexual activity or aggression when compared to their pre-intervention behavior. Chemically sterilized dogs showed a statistically significant increase in dog-directed aggression, but no change in sexual activity. There was no change in home range size in any groups between the pre- and post-intervention measurement. We found no consistent association between levels of serum testosterone concentration and behavioral changes in any of the groups. This study presents the first detailed behavioral observations following surgical and chemical sterilization in male FRDs. The information generated is highly relevant to communities struggling with the control of FRDs. Complementary studies to further our understanding of the effects of male sterilization on the behavioral and reproductive dynamics of FRD populations are needed.

Changes in blood testosterone concentrations after surgical and chemical sterilization of male free-roaming dogs in southern Chile.

There is a growing interest in chemical sterilization as an alternative to surgical castration in large-scale sterilization campaigns to control canine populations. An important step toward understanding the short-term and long-term effects of chemical sterilants is to determine their impact on blood testosterone concentrations, particularly as these could influence dog behavior after treatment. A field trial was conducted with 118 free-roaming male dogs in the Chilean Patagonia, where 36 dogs were chemically sterilized using EsterilSol, 39 dogs were surgically castrated, and 43 dogs remained intact as controls. Blood testosterone levels were determined at four time periods: on enrollment 6 months before treatment (t-6m), at the time of treatment (t0, within one hour after surgical castration or chemical sterilization and during a concurrent 2-week period for the control group), four (t+4m), and six (t+6m) months after treatment. Intrinsic and temporal factors were evaluated; age was significantly associated with testosterone, where dogs 2- to 4-year-old had the highest testosterone concentrations (P = 0.036), whereas body weight and body condition scores were not associated with testosterone; testosterone concentration was not influenced by time of day, month, or season. After treatment (t+4m and t+6m), all of the surgically castrated dogs had testosterone concentrations below 1.0 ng/mL. On the basis of this cut point (<1 ng/mL), testosterone remained unchanged in 66% of the chemically sterilized dogs at both t+4m and t+6m; it remained low for 22% of dogs at both t+4m and t+6m; it was unchanged at t+4m but low at t+6m in 9% of dogs; and, it was low at t+4m but reverted back to unchanged at t+6m in one dog (3%). Incidentally, testosterone in chemically sterilized dogs increased dramatically within 1 hour of treatment (t0), more than doubling (131%) the concentration of control dogs at the time of treatment (t0), likely because of severe necrosis of interstitial cells. The use of EsterilSol as a method of sterilizing dogs had a variable effect on blood testosterone concentrations. Approximately, 30% of chemically sterilized dogs had a reduced testosterone concentration (actual maximum, 1 ng/mL) after 6 months, similar to that of surgically castrated dogs. Most chemically sterilized dogs, however, showed no long-term changes in blood testosterone concentrations.

Necrosuppurative orchitis and scrotal necrotizing dermatitis following intratesticular administration of zinc gluconate neutralized with arginine (EsterilSol) in 2 mixed-breed dogs.

Intratesticular injection of EsterilSol (zinc gluconate neutralized with arginine) is a chemical sterilant for male dogs sometimes used in population control campaigns. Adverse reactions have been reported in 1% to 4% of treated dogs, but detailed histomorphologic descriptions are lacking. During a behavioral study conducted in the Chilean Patagonia in 2012, severe necrosuppurative orchitis and ulcerative dermatitis were observed in 2 of 36 (6%) dogs sterilized with EsterilSol according to the manufacturer's instructions. Reactions were noted on days 8 and 7 postinjection and required scrotal ablation on days 8 and 13, respectively; neither reaction was associated with the injection site. Although self-trauma following administration may have contributed, the cause of the adverse reactions is uncertain. EsterilSol is a relatively uncomplicated method to sterilize male dogs, but the occurrence of severe adverse reactions several days after administration emphasizes the need for the provision of long-term monitoring and veterinary care during sterilization campaigns using this product.

Replication of 2 subtypes of low-pathogenicity avian influenza virus of duck and gull origins in experimentally infected Mallard ducks.

Many subtypes of low-pathogenicity avian influenza (LPAI) virus circulate in wild bird reservoirs, but their prevalence may vary among species. We aimed to compare by real-time reverse-transcriptase polymerase chain reaction, virus isolation, histology, and immunohistochemistry the distribution and pathogenicity of 2 such subtypes of markedly different origins in Mallard ducks (Anas platyrhynchos): H2N3 isolated from a Mallard duck and H13N6 isolated from a Ring-billed Gull (Larus delawarensis). Following intratracheal and intraesophageal inoculation, neither virus caused detectable clinical signs, although H2N3 virus infection was associated with a significantly decreased body weight gain during the period of virus shedding. Both viruses replicated in the lungs and air sacs until approximately day 3 after inoculation and were associated with a locally extensive interstitial, exudative, and proliferative pneumonia. Subtype H2N3, but not subtype H13N6, went on to infect the epithelia of the intestinal mucosa and cloacal bursa, where it replicated without causing lesions until approximately day 5 after inoculation. Larger quantities of subtype H2N3 virus were detected in cloacal swabs than in pharyngeal swabs. The possible clinical significance of LPAI virus-associated pulmonary lesions and intestinal tract infection in ducks deserves further evaluation.

A multicentre prevalence study in Europe on Giardia duodenalis in calves, with molecular identification and risk factor analysis.

The present study aimed to obtain data on the occurrence of Giardia duodenalis in calves in four major cattle rearing countries in Europe (Germany, UK, France and Italy), along with genotyping data and risk factors associated with these infections. A total of 2072 calves were sampled on 207 farms. The majority of the animals were Holstein dairy or mixed Holstein calves (n=1565 or 75.5%), and were female (n=1640 or 79.1%). The average age was 7.8 weeks (SD=4.1; median=7; range=2-16 weeks). All fecal samples were tested using a commercially available monoclonal antibody-based ELISA. The overall apparent prevalence of G. duodenalis for the four countries was 45.4% (n=942/2072) and the overall farm prevalence was 89.9% (186/207), with differences in both animal and farm prevalence between the four countries. The prevalence was significantly higher in animals up to 8 weeks (OR=1.88; P<0.001) compared to older calves, and several management factors including contact with the Dam, Frequency of cleaning of the Maternity Pens, and Disinfection of the Calf Housing were found to be associated with infection. Positive samples were withheld for genotyping using the ß-giardin and triose phosphate isomerase gene: G. duodenalis assemblage E was most prevalent, although 43% of the isolates were typed as assemblage A, with differences in between countries. Furthermore, 32% of the examined samples was found to be a mixed assemblage A and E infection, which is consistent with previous reports. The results of the present study confirm previous findings in other European countries that G. duodenalis infections are common in calves. The infection especially occurs in animals younger than 2 months, and the proportion of positive animals gradually decreased with increasing age.

Prevalence of Batrachochytrium dendrobatidis in three species of wild frogs on Prince Edward Island, Canada.

Chytridiomycosis, caused by the fungus Batrachochytrium dendrobatidis (Bd), has resulted in the decline or extinction of approximately 200 frog species worldwide. It has been reported throughout much of North America, but its presence on Prince Edward Island (PEI), on the eastern coast of Canada, was unknown. To determine the presence and prevalence of Bd on PEI, skin swabs were collected from 115 frogs from 18 separate sites across the province during the summer of 2009. The swabs were tested through single round end-point PCR for the presence of Bd DNA. Thirty-one frogs were positive, including 25/93 (27%) green frogs Lithobates (Rana) clamitans, 5/20 (25%) northern leopard frogs L. (R.) pipiens, and 1/2 (50%) wood frogs L. sylvaticus (formerly R. sylvatica); 12 of the 18 (67%) sites had at least 1 positive frog. The overall prevalence of Bd infection was estimated at 26.9% (7.2-46.7%, 95% CI). Prevalence amongst green frogs and leopard frogs was similar, but green frogs had a stronger PCR signal when compared to leopard frogs, regardless of age (p < 0.001) and body length (p = 0.476). Amongst green frogs, juveniles were more frequently positive than adults (p = 0.001). Green frogs may be the most reliable species to sample when looking for Bd in eastern North America. The 1 wood frog positive for Bd was found dead from chytridiomycosis; none of the other frogs that were positive for Bd by PCR showed any obvious signs of illness. Further monitoring will be required to determine what effect Bd infection has on amphibian population health on PEI.

Primary cerebral hemangiosarcoma in a 6-week-old dog.

A primary cerebral hemangiosarcoma was identified in a 6-week-old, female, cross-breed dog. Grossly, the tumor mass was poorly demarcated from the adjacent neuropil, hemorrhagic, and caused effacement of the right dorsolateral cerebral hemisphere. Microscopically, the tumor was composed of an infiltrative mass of small vascular channels lined by neoplastic endothelial cells that stained variably with factor VIII-related antigen and negatively with glial fibrillary acidic protein. This is the first description of a primary intracranial hemangiosarcoma in an immature dog.