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D H Graham

Publications and source records attributed to D H Graham.

At least 19 recordsLinked to original sources

Molecular and biologic characteristics of Toxoplasma gondii isolates from wildlife in the United States.

Toxoplasma gondii isolates can be grouped into 3 genetic lineages. Type I isolates are considered more virulent in outbred mice and have been isolated predominantly from clinical cases of human toxoplasmosis, whereas types II and III isolates are considered less virulent for mice and are found in humans and food animals. Little is known of genotypes of T. gondii isolates from wild animals. In the present report, genotypes of isolates of T. gondii from wildlife in the United States are described. Sera from wildlife were tested for antibodies to T. gondii with the modified agglutination test, and tissues from animals with titers of 1:25 (seropositive) were bioassayed in mice. Toxoplasma gondii was isolated from the hearts of 21 of 34 seropositive white-tailed deer (Odocoileus virginianus) from Mississippi and from 7 of 29 raccoons (Procyon lotor); 5 of 6 bobcats (Lynx rufus); and the gray fox (Urocyon cinereoargenteus), red fox (Vulpes vulpes), and coyote (Canis latrans) from Georgia. Toxoplasma gondii was also isolated from 7 of 10 seropositive black bears (Ursus americanus) from Pennsylvania by bioassay in cats. All 3 genotypes of T. gondii based on the SAG2 locus were circulating among wildlife.

Animals↗

Genotyping of Toxoplasma gondii isolates from chickens from India.

The present study was undertaken to isolate and genotype Toxoplasma gondii from free-range chickens (Gallus domesticus) from villages in Maharashtra and Tamil Nadu states of central and south India, respectively. Blood, heart, and brain from a total of 741 chickens were examined for T. gondii infection. Antibodies to T. gondii, as assayed with the modified agglutination test (MAT >or = 1:5) were found in 133 (17.9%) chickens. Hearts and brains of 186 chickens were bioassayed in mice. Additionally, hearts and/or brains of most of the seronegative (MAT < 1:5) chickens were fed to 20 T. gondii-free cats, while 32 seropositive chickens (MAT 1:5) were fed to 3 cats. T. gondii was not isolated from any of the chickens by mouse bioassay. Five of the cats that were fed seronegative chickens shed oocysts, while isolates were not obtained from any of the other cats fed seropositive chickens. These five isolates, along with the two that were previously isolated in India through cat bioassay, were genetically analyzed. Genotyping using the SAG 2 locus indicated that two isolates were type II and five were type III. Microsatellite analysis revealed allelic differences between and within the lineages. This is the first report of genetic characterization of any T. gondii isolate from India.

Agglutination Tests↗

Characterization of Toxoplasma gondii isolates from free range chickens from Paraná, Brazil.

The prevalence of Toxoplasma gondii in free range chickens is a good indicator of the prevalence of T. gondii oocysts in the environment because chickens feed from the ground. In the present study, prevalence of T. gondii in 40 free range chickens (Gallus domesticus) from a rural area surrounding Paraná, Brazil was assessed. Blood, heart, and brain from each chicken were examined for T. gondii infection. Antibodies to T. gondii, assayed with the modified agglutination test (MAT> or =1:5) were found in 16 chickens. Hearts and brains of seropositive (MAT> or =1:5) chickens were bioassayed in mice. Additionally, hearts and brains of seronegative (MAT<1:5) chickens were bioassayed in two T. gondii-free cats (12 chickens per cat). T. gondii was isolated from 13 of 16 (81%) seropositive chickens. Of the two cats fed tissues pooled form seronegative chickens, one shed T. gondii oocysts. Nine of the 13 T. gondii isolates killed 100% of infected mice. The T. gondii isolate from the cat was also virulent for mice. Genotyping of 13 chicken isolates of T. gondii using the SAG2 locus indicated that seven isolates were type I and six were type III; three of these type III isolates killed all infected mice suggesting that all strains virulent for mice are not type I. The isolate from the feces of the cat fed chicken tissues was type I.

Agglutination Tests↗

Isolation and molecular characterization of Toxoplasma gondii from chickens and ducks from Egypt.

The prevalence of Toxoplasma gondii in free range chickens is a good indicator of the prevalence of T. gondii oocysts in the environment because chickens feed from the ground. In the present study, prevalence of T. gondii in 121 free range chickens (Gallus domesticus) and 19 ducks (Anas sp.) from a rural area surrounding Giza, Egypt was assessed. Blood, heart, and brain from each animal were examined for T. gondii infection. Antibodies to T. gondii, assayed with the modified agglutination test (MAT), were found in 49 (40.4%) chickens in titers of 1:5 in 11, 1:10 in four, 1:20 in four, 1:40 in eight, 1:80 in 10, and 1:160 or more in 12 chickens. Antibodies were found in three ducks each with a titer of 1:80. Hearts and brains of seropositive (MAT > or = 1:5) chickens and ducks were bioassayed in mice. Additionally, hearts and brains of seronegative (MAT<1:5) animals were bioassayed in T. gondii-free cats. T. gondii was isolated from 19 of 49 seropositive chickens (one with a titer of 1:5, two with a titer of 1:20, one with a titer of 1:40, five with a titer of 1:80, three with a titer of 1:160, and seven with a titer of > or = 1:360). One cat fed tissues pooled from 15 seronegative chickens shed T. gondii oocysts, while two cats fed tissues of 34 seronegative chickens did not shed oocysts. T. gondii was isolated from one of the seropositive ducks by bioassay in mice. The two cats fed tissues from 16 seronegative ducks did not shed oocysts. Genotyping of 20 chicken isolates of T. gondii using the SAG 2 locus indicated that 17 isolates were type III and three were type II. The duck isolate of T. gondii was type III. The mice inoculated with tissue stages of all 21 isolates of T. gondii from chickens and ducks remained asymptomatic, indicating that phenotypically they were not type I because type I strains are lethal for mice. Infections with mixed genotypes were not found.

Agglutination Tests↗

Toxoplasma gondii isolates from free-ranging chickens from the United States.

Prevalence of Toxoplasma gondii infection in chickens is a good indicator of the strains prevalent in their environment because they feed from ground. The prevalence of T. gondii was determined in 118 free-range chickens from 14 counties in Ohio and in 11 chickens from a pig farm in Massachusetts. Toxoplasma gondii antibodies (> or = 1: 5) were found using the modified agglutination test (MAT) in 20 of 118 chickens from Ohio. Viable T. gondii was recovered from 11 of 20 seropositive chickens by bioassay of their hearts and brains into mice. The parasite was not isolated from tissues of 63 seronegative (< or = 1:5) chickens by bioassay in cats. Hearts, brains, and muscles from legs and breast of the 11 chickens from the pig farm in Massachusetts were fed each to a T. gondii-negative cat. Eight cats fed chicken tissues shed oocysts; the 3 cats that did not shed oocysts were fed tissues of chickens with MAT titers of 1:5 or less. Tachyzoites of 19 isolates of T. gondii from Ohio and Massachusetts were considered avirulent for mice. Of 19 isolates genotyped, 5 isolates were type II and 14 were type III; mixed types and type I isolates were not found.

Agglutination Tests↗

Isolation and genotyping of Toxoplasma gondii from free-ranging chickens from Argentina.

The prevalence of Toxoplasma gondii in free-ranging chickens can be considered a good indicator of the prevalence of T. gondii oocysts in the environment because chickens feed from the ground. In the present study, prevalence of T. gondii in 29 free-range chickens (Gallus domesticus) from Argentina was investigated. Blood, heart, and brain from each chicken were examined for T. gondii infection. Antibodies to T. gondii, assayed with the modified agglutination test (MAT), were found in 19 of 29 (65.5%) chickens. Hearts and brains of seropositive (MAT > or = 1:5) chickens were bioassayed in mice. Toxoplasma gondii was isolated from 9 of 19 seropositive chickens. Genotyping of chicken isolates of T. gondii using the SAG2 locus indicated that 1 was type I, 1 was type II, and 7 were type III. This is the first report of isolation of T. gondii from chickens from Argentina.

Agglutination Tests↗

Toxoplasma gondii isolates of free-ranging chickens from Rio de Janeiro, Brazil: mouse mortality, genotype, and oocyst shedding by cats.

Most isolates of Toxoplasma gondii can be grouped into 3 genetic lineages. In the present study, 67 isolates of T. gondii were obtained by bioassay in mice inoculated with brains and hearts of 96 asymptomatic chickens from an area highly endemic to human infection in Rio de Janeiro, Brazil. Of the 48 isolates genotyped using the SAG2 locus, 34 (70%) were of type I and 13 (27%) were of type III. No isolate of type II was recovered. Isolates from 1 chicken contained a type I and type III mixed infection, indicating natural multiparasite infection in the same animal. Cats fed mice infected with 11 type I strains shed 19-535 million oocysts in their feces, indicating that type I isolates can circulate in the environment.

Animals↗

Biological and genetic characterisation of Toxoplasma gondii isolates from chickens (Gallus domesticus) from São Paulo, Brazil: unexpected findings.

In spite of a wide host range and a world wide distribution, Toxoplasma gondii has a low genetic diversity. Most isolates of T. gondii can be grouped into two to three lineages. Type I strains are considered highly virulent in outbred laboratory mice, and have been isolated predominantly from clinical cases of human toxoplasmosis whereas types II and III strains are considered avirulent for mice. In the present study, 17 of 25 of the T. gondii isolates obtained from asymptomatic chickens from rural areas surrounding São Paulo, Brazil were type I. Antibodies to T. gondii were measured in 82 chicken sera by the modified agglutination test using whole formalin-preserved tachyzoites and mercaptoethanol and titres of 1:10 or more were found in 32 chickens. Twenty-two isolates of T. gondii were obtained by bioassay in mice inoculated with brains and hearts of 29 seropositive (> or =1:40) chickens and three isolates were obtained from the faeces of cats fed tissues from 52 chickens with no or low levels (<1:40) of antibodies. In total, 25 isolates of T. gondii were obtained by bioassay of 82 chicken tissues into mice and cats. All type I isolates killed all infected mice within 4 weeks whereas type III isolates were less virulent to mice. There were no type II strains. Tissue cysts were found in mice infected with all 25 isolates and all nine type I isolates produced oocysts. Infected chickens were from localities that were 18-200 km apart, indicating no common source for T. gondii isolates. This is the first report of isolation of predominantly type I strains of T. gondii from a food animal. Epidemiological implications of these findings are discussed.

Agglutination Tests↗

Documenting patient refusals.

Patient morbidity and mortality, subsequent to either patient- or provider-initiated refusals, are noteworthy. It has been estimated that hospital admission is twice as likely after prehospital providers refuse a patient transportation to the hospital. In one particular study group, prehospital-provider refusal of transportation, as opposed to patient refusal of transportation, accounted for 73% of the post-refusal hospital admissions. Provider-initiated refusals are tantamount to a time bomb. There are very few justifiable provider-initiated refusals of treatment or transportation. If the call is of a non-emergency nature, the decision not to treat or transport should be a mutual agreement between the patient and provider. Consult with the online medical director for guidance as needed. Document the physician's name, consulting facility and medical direction. As with every patient encounter, a legally defensible runsheet should be completed. Should you write a report if your services are "not needed," or if you are canceled en route to the call? For your protection, a report archiving every run should be documented. If your services are canceled en route, note the canceling authority and time of cancellation. If your services are canceled at the scene, document the canceling authority, time of cancellation and the circumstance. It is important to specifically document that "no patient contacts were made." When patient contact is made, a patient-provider relationship is established, thereby redefining your duty to the patient. Protect yourself, your crew members, your chain of command, your jurisdiction and your agency by writing a legally defensible informed refusal report. According to one source, "Every negligence case in the last 30 years has been decided on its documentation." If it wasn't written down, it wasn't done. Be safe, and document safely.

Documentation↗

Selection of refractory and permissive strains of Aedes triseriatus (Diptera: Culicidae) for transovarial transmission of La Crosse virus.

The genetic basis of transovarial transmission of La Crosse virus in Aedes triseriatus (Say) was investigated through selection experiments on 2 mosquito strains. One strain was subject to selection for transovarial transmission refractoriness, the other for permissiveness to transovarial transmission. Response to selection for a low filial infection rate was rapid, decreasing from 18 to 3% in 3 generations. However, no response to selection for permissiveness was observed in the other strain; the average filial infection rates through 4 generations fluctuated between 25 and 40%. By contrast, the transovarial transmission rate in both strains showed a consistent response to selection in both directions. These patterns are consistent with a model in which transovarial transmission is controlled by a single genetic locus and permissiveness is conditioned by dominant alleles; whereas the filial infection rate is nongenetic and influenced by stochastic factors in the mosquito and virus.

Aedes↗