Isolation and Annotation of the Genome Sequences of Bacteriophages InvictusManeo (Subcluster K5) and Netyap (Subcluster L2).

The mycobacteriophages InvictusManeo (K5 subcluster) and Netyap (L2 subcluster) were isolated from soils in Cullowhee Creek, Cullowhee, North Carolina. Both exhibit Siphoviridae morphology and infect Mycobacterium smegmatis mc2155. The InvictusManeo genome is 61,147 bp and contains 96 predicted protein-coding genes, whereas the Netyap genome is 76,366 bp with 131 predicted protein-coding genes.

Obtaining an isolate of Ancylostoma braziliense from cats without the need for necropsy.

Because the eggs of Ancylostoma species of dogs and cats are difficult to readily distinguish morphologically, isolation of a certain species often requires the humane death of the source animal or holding an animal after treatment to obtain worms for specific identification or to harvest ex utero eggs. The objective of this study was to obtain an isolate of Ancylostoma braziliense from 1-time, field-collected samples of feline feces without the need for the killing of any animals. During a collection trip to Florida, fecal samples (n  =  40) were collected and identified as containing A. braziliense eggs (n  =  26) using centrifugal sugar flotation. Eggs from hookworm-positive slides were washed into tubes, DNA was extracted, and 10 samples were identified as containing A. braziliense using restriction fragment length polymorphism (RFLP) with Hinf1. Six of these samples also contained DNA of Ancylostoma tubaeforme and, thus, only 4 samples were from cats infected only with A. braziliense. Larvae cultured from two of the latter samples were used to subcutaneously inoculate a purpose-bred puppy with the intention to inhibit the growth of any potentially contaminating A. tubaeforme larvae in the culture. The infection was patent at 14 days after inoculation, and the eggs were identified as A. braziliense by RFLP and DNA sequencing. Larvae were cultured from the feces of this dog and used to infect a laboratory-reared, specific-pathogen-free cat; the eggs and larvae produced by the cat were also identified molecularly as those of A. braziliense. The larvae from this cat were used to infect other cats to maintain the isolate for further research. Both the puppy and the first cat used in this study were treated to clear their infections and have since been adopted by new owners.

Treatment of Baylisascaris procyonis infections in dogs with milbemycin oxime.

An examination was made as to the ability of Sentinel Flavor Tabs (milbemycin oxime/lufenuron) to treat Baylisascaris procyonis infections in dogs. The study was designed as a critical trial and included five naturally infected dogs and two dogs that were experimentally infected. Another dog from a prior clinical trial that was treated with Sentinel Flavor Tabs as part of the original FDA submission package for intestinal nematode infections was also included with the treated dogs. Of the five naturally infected dogs treated as part of the critical trial, three were cleared of their infections. These five dogs passed a total of 52 worms after treatment; one dog retained 23 worms and the other retained 1 worm at necropsy 7 days after treatment. Two of five experimentally infected Beagle dogs that had been given mice that had been fed 200 infectious eggs, developed patent infections with the parasite. These dogs were treated, and one of the dogs passed one worm and the other passed two worms after treatment with no worms being detected at necropsy 7 days after treatment. The one dog that was treated with milbemycin oxime as part of the FDA submission was clear of worms at necropsy. Overall, the mean efficacy of Sentinel Flavor Tabs was found to be 91.0%. Of the eight dogs that were treated, six were totally cleared of their infections, a cure rate of 75%. The two dogs that did not clear their infections had very large numbers of adult B. procyonis within their intestinal tracts at the time of treatment, one dog had 40 worms (23 remaining) and the other had 26 worms (1 remaining). It is suggested that the treatment of dogs with monthly Sentinel Flavor Tabs could markedly reduce the chance of infected dogs contaminating the environment. Also, additional monthly treatments are highly likely to clear dogs of any worms not killed with the initial treatment.