Melioidosis in Cynomolgus Macaques ( Macaca Fascicularis ) Imported to the United States from Cambodia.

Melioidosis, a potentially fatal infectious disease of humans and animals, including nonhuman primates (NHPs), is caused by the high-consequence pathogen Burkholderia pseudomallei. This environmental bacterium is found in the soil and water of tropical regions, such as Southeast Asia, where melioidosis is endemic. The global movement of humans and animals can introduce B. pseudomallei into nonendemic regions of the United States, where environmental conditions could allow establishment of the organism. Approximately 60% of NHPs imported into the United States originate in countries considered endemic for melioidosis. To prevent the introduction of infectious agents to the United States, the Centers for Disease Control and Prevention (CDC) requires newly imported NHPs to be quarantined for at least 31 d, during which time their health is closely monitored. Most diseases of public health concern that are transmissible from imported NHPs have relatively short incubation periods that fall within the 31-d quarantine period. However, animals infected with B. pseudomallei may appear healthy for months to years before showing signs of illness, during which time they can shed the organism into the environment. Melioidosis presents diagnostic challenges because it causes nonspecific clinical signs, serologic screening can produce unreliable results, and culture isolates are often misidentified on rapid commercial testing systems. Here, we present a case of melioidosis in a cynomolgus macaque (Macaca fascicularis) that developed a subcutaneous abscess after importation from Cambodia to the United States. The bacterial isolate from the abscess was initially misidentified on a commercial test. This case emphasizes the possibility of melioidosis in NHPs imported from endemic countries and its associated diagnostic challenges. If melioidosis is suspected, diagnostic samples and culture isolates should be submitted to a laboratory in the CDC Laboratory Response Network for conclusive identification and characterization of the pathogen.

Public Health Implications of Changing Rodent Importation Patterns - United States, 1999-2013.

The United States imports a large volume of live wild and domestic animal species; these animals pose a demonstrated risk for introduction of zoonotic diseases. Rodents are imported for multiple purposes, including scientific research, zoo exhibits and the pet trade. Current U.S. public health regulatory restrictions specific to rodent importation pertain only to those of African origin. To understand the impacts of these regulations and the potential public health risks of international rodent trade to the United States, we evaluated live rodent import records during 1999-2013 by shipment volume and geographic origin, source (e.g. wild-caught versus captive- or commercially bred), intended purpose and rodent taxonomy. Live rodent imports increased from 2737 animals during 1999 to 173 761 animals during 2013. Increases in both the number and size of shipments contributed to this trend. The proportion of wild-captured imports declined from 75% during 1999 to <1% during 2013. Nearly all shipments during these years were imported for commercial purposes. Imports from Europe and other countries in North America experienced notable increases in volume. Gerbils and hamsters arriving from Europe and chinchillas, guinea pigs and hamsters arriving from other countries in North America were predominant taxa underlying this trend. After 2003, African-origin imports became sporadic events under the federal permit process. These patterns suggest development of large-scale captive rodent breeding markets abroad for commercial sale in the United States. While the shift from wild-captured imports alleviates many conservation concerns and risks for novel disease emergence, such consolidated sourcing might elevate exposure risks for zoonotic diseases associated with high-density rodent breeding (e.g. lymphocytic choriomeningitis or salmonellosis). A responsive border health system must periodically re-evaluate importation regulations in conjunction with key stakeholders to ensure a balance between the economic benefits of rodent trade against the potential public health risks.

Use of nonhuman primates in research in North America.

In North America, the biomedical research community faces social and economic challenges to nonhuman primate (NHP) importation that could reduce the number of NHP available for research needs. The effect of such limitations on specific biomedical research topics is unknown. The Association of Primate Veterinarians (APV), with assistance from the Centers for Disease Control and Prevention, developed a survey regarding biomedical research involving NHP in the United States and Canada. The survey sought to determine the number and species of NHP maintained at APV members' facilities, current uses of NHP to identify the types of biomedical research that rely on imported animals, and members' perceived trends in NHP research. Of the 149 members contacted, 33 (22%) replied, representing diverse facility sizes and types. Cynomolgus and rhesus macaques were the most common species housed at responding institutions and comprised the majority of newly acquired and imported NHP. The most common uses for NHP included pharmaceutical research and development and neuroscience, neurology, or neuromuscular disease research. Preclinical safety testing and cancer research projects usually involved imported NHP, whereas research on aging or degenerative disease, reproduction or reproductive disease, and organ or tissue transplantation typically used domestic-bred NHP. The current results improve our understanding of the research uses for imported NHP in North America and may facilitate estimating the potential effect of any future changes in NHP accessibility for research purposes. Ensuring that sufficient NHP are available for critical biomedical research remains a pressing concern for the biomedical research community in North America.

Is that a rodent in your luggage? A mixed method approach to describe bushmeat importation into the United States.

Bushmeat, defined as meat derived from wild animals, is a potential source of zoonotic pathogens. Bushmeat from restricted animals is illegal to import into the United States under US federal regulations. We reviewed US Centers for Disease Control and Prevention (CDC) port of entry surveillance records from September 2005 through December 2010 and conducted focus group studies to describe trends in and reasons for bushmeat importation into the United States. In total, 543 confiscated bushmeat items were recorded. Half of the confiscated bushmeat items identified were rodents. Africa was the most frequent continent of origin. Seasonality was evident, with bushmeat confiscations peaking in late spring to early summer. Four times more bushmeat was confiscated during an enhanced surveillance period in June 2010 compared with the same period in previous years, suggesting that routine surveillance underestimated the amount of bushmeat detected at US Ports of Entry. Focus groups held in three major US cities revealed that bushmeat importation is a multifaceted issue. Longstanding cultural practices of hunting and eating bushmeat make it difficult for consumers to acknowledge potential health and ecologic risks. Also, US merchants selling African goods, including bushmeat, in their stores have caused confusion among importers as to whether importation is truly illegal. Enhancing routine surveillance for bushmeat and consistent enforcement of penalties at all ports of entry, along with health education aimed at bushmeat importers, might be useful to deter illegal importation.

Prevention and control of rabies in an age of global travel: a review of travel- and trade-associated rabies events--United States, 1986-2012.

Rabies prevention and control efforts have been successful in reducing or eliminating virus circulation regionally through vaccination of specific reservoir populations. A notable example of this success is the elimination of canine rabies virus variant from the United States and many other countries. However, increased international travel and trade can pose risks for rapid, long-distance movements of ill or infected persons or animals. Such travel and trade can result in human exposures to rabies virus during travel or transit and could contribute to the re-introduction of canine rabies variant or transmission of other viral variants among animal host populations. We present a review of travel- and trade-associated rabies events that highlight international public health obligations and collaborative opportunities for rabies prevention and control in an age of global travel. Rabies is a fatal disease that warrants proactive coordination among international public health and travel industry partners (such as travel agents, tour companies and airlines) to protect human lives and to prevent the movement of viral variants among host populations.

Mosquito infection studies with Aotus monkeys and humans infected with the Chesson strain of Plasmodiun vivax.

Oocyst counts were compared between mosquitoes that fed on humans versus mosquitoes that fed on Aotus monkeys, both of which were infected with the Chesson strain of Plasmodium vivax. Oocyst counts obtained from mosquitoes fed on humans were almost 10-fold higher in number. Mosquitoes were more likely to be infected and with a higher rate of infection when they fed on monkeys before the peak in the asexual parasite count. Mosquitoes that fed on humans were more likely to be more heavily infected when fed after the peak in the asexual count. Of several species of owl monkeys, Aotus vociferans was infected at a higher frequency. On the basis of oocyst counts, Anopheles dirus were the most susceptible and An. maculatus were the least susceptible of the mosquito species tested.

Zoonotic viruses associated with illegally imported wildlife products.

The global trade in wildlife has historically contributed to the emergence and spread of infectious diseases. The United States is the world's largest importer of wildlife and wildlife products, yet minimal pathogen surveillance has precluded assessment of the health risks posed by this practice. This report details the findings of a pilot project to establish surveillance methodology for zoonotic agents in confiscated wildlife products. Initial findings from samples collected at several international airports identified parts originating from nonhuman primate (NHP) and rodent species, including baboon, chimpanzee, mangabey, guenon, green monkey, cane rat and rat. Pathogen screening identified retroviruses (simian foamy virus) and/or herpesviruses (cytomegalovirus and lymphocryptovirus) in the NHP samples. These results are the first demonstration that illegal bushmeat importation into the United States could act as a conduit for pathogen spread, and suggest that implementation of disease surveillance of the wildlife trade will help facilitate prevention of disease emergence.

Infection of mosquitoes with Plasmodium falciparum by feeding on humans and on Aotus monkeys.

Of 1,004 positive lots of mosquitoes fed on 229 humans infected with Plasmodium falciparum, 46.2% had 1-10 oocysts/(+)gut, 21.2% had 10-30 oocysts/(+)gut, 22.2% had 30-100 oocysts/(+)gut, and 10.4% had > 100 oocysts/(+) gut. The highest levels of infection occurred between 6 and 15 days after the peak in the asexual parasite count. Of 2,281 lots of Anopheles freeborni mosquitoes fed on splenectomized Aotus monkeys infected with the Santa Lucia strain of P. falciparum, 1,191 were infected (52.2%). The highest intensity infections ranged from 2.78 oocysts per positive gut in mosquitoes fed on Aotus vociferans to 6.08 oocysts per positive gut for those fed on A. lemurinus griseimembra to 10.4 oocysts per positive gut for those fed on A. nancymaae. The pattern of infection for mosquitoes fed on splenectomized Aotus monkeys was similar to that obtained by feeding on humans, but the intensity, based on oocyst/(+)gut, was much lower.

Plasmodium fieldi: observations on the Hackeri and ABI strains in Macaca mulatta monkeys and mosquitoes.

Macaca mulatta monkeys infected with the Hackeri strain of Plasmodium fieldi had maximum parasite counts ranging from 1,300 to 301,320/microL. In 43 intact animals infected with the ABI strain, the maximum parasite counts ranged from 672 to 57,189/microL (median = 15,100/microL); in 46 splenectomized monkeys, the maximum parasite count ranged from 660 to 350,000/microL (median = 52,245/microL). Transmission through Anopheles dirus mosquitoes was obtained on 11 occasions with pre-patent periods of 9-14 days. Relapses occurred between two and eight times during a 1-year period. P. fieldi has potential for testing prophylactic and radical curative drugs.

The Santa Lucia strain of Plasmodium falciparum in Aotus monkeys.

The Santa Lucia strain of Plasmodium falciparum was studied in 150 Aotus lemurinus griseimembra, 30 A. azarae boliviensis, 103 A. nancymaae, and 121 A. vociferans monkeys. All four of these splenectomized hosts supported the production of gametocytes infective to Anopheles freeborni mosquitoes. Transmission through sporozoites from An. freeborni, An. stephensi, An. maculatus, and An. albimanus mosquitoes was successful to all four species of Aotus on a total of 100 occasions with a median pre-patent period of 21 days. For the production of infective mosquitoes for vaccine challenge studies, A. l. griseimembra and A. vociferans were the most predictable hosts.

Border health: who's guarding the gate?

Changes in the global trade market have led to a thriving international pet trade in exotic animals, birds, and puppies. The flood of animals crossing the United States' borders satisfies the public demand for these pets but is not without risk. Imported pets may be infected with diseases that put animals or the public at risk. Numerous agencies work together to reduce the risk of animal disease introduction, but regulations may need to be modified to ensure compliance. With more than 280,000 dogs and 183,000 wildlife shipments being imported into the United States each year, veterinarians must remain vigilant so they can recognize potential threats quickly.

Studies on the Salvador I strain of Plasmodium vivax in non-human primates and anopheline mosquitoes.

A review is presented on studies conducted in New World monkeys and chimpanzees with the Salvador I strain of Plasmodium vivax. This isolate has been adapted to Aotus and Saimiri (squirrel) monkeys and developed as a model for the testing of antimalarial vaccines. After the injection of 10,000 sporozoites, the median prepatent period in S. boliviensis monkeys was 21.5 days. In 103 sporozoite-induced infections in splenectomized monkeys, the median maximum parasite count ranged from 2,139 to 202,368/microL, with a median maximum parasite count of 48,174/microL. Median maximum parasite counts in Aotus lemurinus griseimembra, A. nancymaae, A. azarae boliviensis, and A. vociferans monkeys were 19,902, 18,390, 21,420, and 18,210/microL, respectively and ranged from 124 to 156,000/microL. Mosquito infections were readily obtained in different species of Anopheles mosquitoes. The S. boliviensis monkey and Salvador I strain seems suitable for the testing of sporozoite and liver stage vaccines but not for blood-stage vaccines against P. vivax unless adapted further in spleen-intact Saimiri boliviensis monkeys.

The Chesson strain of plasmodium vivax in humans and different species of Aotus monkeys.

Comparison was made between the parasitemia of Chesson strain Plasmodium vivax in humans and in splenectomized Aotus lemurinus griseimembra, A. nancymaae, A. vociferans, and A. azarae boliviensis monkeys. In the monkeys, 56.3% of the animals had maximum counts > 25,000/muL and in humans 59.6% were above this peak parasitemia. In humans, it took an average of 9.3 days to reach the maximum parasite count. In monkeys with no previous infections, it took an average of 18.9 days to reach the maximum parasite count; for those with previous infections, it took an average of 15 days. Human and nonhuman primate data on this parasite suggest that splenectomized Aotus monkeys, particularly A. lemurinus griseimembra, and to a somewhat lesser extent A. vociferans, can mimic the course of Chesson malaria in humans regarding parasitemia and mosquito infection.

Importation of dogs into the United States: risks from rabies and other zoonotic diseases.

The importation of dogs into the United States poses a risk for the introduction of rabies and other zoonotic diseases. Federal regulations (42 CFR 71.51) currently require proof of valid rabies vaccination for imported dogs, but allow the importation of some unvaccinated dogs, including dogs less than 3 months of age, provided certain requirements for confinement are met until the dog is vaccinated. Although there are no accurate surveillance data on the number of dogs imported each year, it is estimated based on extrapolated data that over 287,000 dogs were imported into the United States during 2006. Of these, approximately 25% were either too young to be vaccinated or lacked proof of valid rabies vaccination. Import trends suggest that an increasing number of unvaccinated puppies are being imported into the United States, many through commercial resale or rescue operations. Since 2004, foreign canine rabies virus variants have been documented in at least two imported puppies. Federal regulations are currently being reviewed by the Centers for Disease Control and Prevention to determine if they can be updated to address current import trends and disease risks, such as requiring a health screen and valid rabies vaccinations for all dogs prior to entry.

Thermal decomposition studies of the polyhedral oligomeric silsesquioxane, POSSh, and when it is impregnated with the metallocene bis(eta5-cyclopentadienyl)zirconium (IV) dichloride or immobilized on silica.

Thermal decomposition studies of the free polyhedral oligomeric silsesquioxane, POSSh, and when this compound has been impregnated with Cp2ZrCl2 (Cp = eta5-C5H5) or immobilized on SiO2 were conducted using infrared emission spectroscopy (IES) over a 100-1000 degrees C temperature range and by thermogravimetric analysis (TGA). The organic groups in POSS(h) apparently decompose thermally into Si-CH3, Si-H and other fragments. Upon impregnation with Cp2ZrCl2, however, a different thermal decomposition pathway was followed and new infrared emission bands appeared in the 1000-900 cm(-1) region suggesting the formation of Si-O-Zr moieties. When immobilized on SiO2 and subjected to thermal decomposition, the POSSh compound lost its organic groups and the inorganic structure remaining was incorporated into the SiO2 framework.

Adaptation of a multi-drug resistant strain of Plasmodium falciparum from Peru to Aotus lemurinus griseimembra, A. nancymaae, and A. vociferans monkeys.

A strain of Plasmodium falciparum from Peru was adapted to splenectomized Aotus nancymaae and Aotus vociferans monkeys. The Peru 134/CDC strain of P. falciparum was shown to be resistant to treatment with chloroquine in monkeys and partially resistant to mefloquine and malarone. Genetic mutations in crt, dhfr, dhps, and cytochrome b genes conferring drug resistance were also determined for this Peruvian strain of P. falciparum.

Vibrational spectra of silsesquioxanes impregnated with the metallocene catalyst bis(eta(5)-cyclopentadienyl)zirconium(IV) dichloride.

FT-IR photoacoustic and Raman spectroscopy have been used to study the interactions between the metallocene catalyst, Cp(2)ZrCl(2) (Cp=eta(5)-C(5)H(5)), and two polyhedral oligomeric silsesquioxanes (POSS) supports. The first silsesquioxane support, POSS(h), contains (beta-hydroxyl)-tertiary amine groups, while in the second one, POSS(u), these -OH groups have been converted into N-(p-toluyl) urethane groups. The vibrational spectra of the Cp(2)ZrCl(2):POSS(h) and Cp(2)ZrCl(2):POSS(u) samples show that the Cp(2)ZrCl(2) catalyst reacts with the C-OH groups of POSS(h) and also interacts with N-H and >CO groups of POSS(u). Furthermore, Cp(2)ZrCl(2) can react with the Si-OH groups of the POSS supports and also interact with the O atoms that are bonded to the benzene rings and the N atoms of the tertiary amines in both silsesquioxanes. As a result of the interactions between Cp(2)ZrCl(2) and the POSS supports, acidic species are generated. The Cp(2)ZrCl(2):POSS(h) mass ratio seems to be an important parameter in the formation of Zr-O bonds and the acidic species.

Isolates of Plasmodium inui adapted to Macaca mulatta monkeys and laboratory-reared anopheline mosquitoes for experimental study.

Plasmodium inui is a parasite of macaques and other nonhuman primates in Asia that is studied as a model for the human malaria parasite P. malariae. Presented here are descriptions of the isolation, passage histories into Macaca mulatta monkeys, and infectivity to different Anopheles spp. mosquitoes of 18 different isolates of this parasite.

Studies on sporozoite-induced and chronic infections with Plasmodium fragile in Macaca mulatta and New World monkeys.

Plasmodium fragile continues to be investigated because of its biologic similarities to the human malaria parasite, Plasmodium falciparum. Two strains of P. fragile are available for study; one strain is able to infect mosquitoes, whereas the other strain is transmissible only by blood inoculation. The Sri Lanka strain of P. fragile was transmitted to Macaca mulatta, Macaca fascicularis, Aotus lemurinus griseimembra, Aotus nancymaae, Aotus vociferans, and Saimiri boliviensis monkeys via sporozoites that developed to maturity only in Anopheles dirus mosquitoes. The prepatent periods ranged from 12 to 35 days for macaques and from 15 to 30 days for New World monkeys after intravenous injection of sporozoites. Eight rhesus monkeys were infected with the Nilgiri strain and followed for 482 days. Parasitemia in 6 animals persisted at relatively high density through the period of observation. Erythrocyte, hematocrit, and hemoglobin values reached their lowest levels 3 wk after infection and slowly recovered; however, the values did not approach preinfection levels as long as parasitemia persisted in the monkeys. The mean corpuscular volume and corpuscular hemoglobin concentration reached their peak and lowest values, respectively, at day 38 and then returned to the preinfection level. The mean corpuscular hemoglobin value decreased to its lowest level at day 87 and then returned to preinfection level.