Enzootic Foci Research Articles

Molecular evolution of eastern equine encephalomyelitis virus in North America

We examined the rate and spatial pattern of eastern equine encephalomyelitis virus (EEEV) evolution in North America using primer-extension sequencing of viral RNA. Nucleotide sequences of the entire 26 S structural gene region of four EEEV strains revealed remarkable conservation between 1933 and 1985, with an estimated 0.7% divergence or 1.4 × 10 −4 nucleotide substitutions per site per year. Sequences from smaller 26 S regions of nine additional strains suggested that EEEV evolves in North America in a single lineage, with genetic exchange regularly occurring among enzootic transmission foci. In these limited 26 S genome regions, only synonymous nucleotide substitutions became fixed between 1933 and 1988, implying a high degree of conservation in protein structure. Short nucleotide sequences from a Panamanian, South American variety isolate revealed a relatively distant relationship to North American serotype viruses. This suggested genetic divergence between antigenic varieties, and independent evolution of EEEV in North and South America. Factors related to replication and epidemiology of EEEV, which may constrain its evolution in nature, are discussed. Possible mechanisms of genetic exchange among enzootic foci are also considered.

Plague and Tularemia

Human plague is a local or systemic flea-transmitted infection caused by Yersinia pestis. It is maintained in well established enzootic foci among wild rodents. This article discusses the clinical findings in plague, diagnosis, treatment, and prevention of plague, and management of contacts of human plague cases and of exposures to epizootic plague. Tularemia shares many features with plague but is widespread in animal and arthropod vector populations and essentially throughout the United States.

Vesicular stomatitis new jersey virus glycoprotein gene sequence and neutralizing epitope stability in an enzootic focus

Vesicular stomatitis New Jersey (VS NJ) virus is capable of undergoing rapid evolution in nature and therefore has the potential for antigenic variation. We selected an area of Costa Rica where VS NJ virus is enzootic to study whether this virus used the mechanism of antigenic variation to persist in nature. Three sentinel herds and three nonsentinel herds were observed from 1986 to 1988. Eleven VS N1 virus isolates were collected from naturally infected cattle. Remarkably, nine animals that were bled prior to reinfection with VS NJ virus had neutralizing antibody titers up to 1: 102,400 yet virus was isolated from, and disease was observed in, these animals. Sequence analysis of the portion of the glycoprotein gene coding for the neutralizing epitopes demonstrated that all virus isolates were 98–100% similar with no indication of specific genetic variation. The 3′ end of the glycoprotein gene also remained stable in that all isolates were again 98–100% similar in nucleotide sequence. Each isolate was neutralized to equivalent titers with monoclonal antibodies directed against four neutralizing epitopes on the glycoprotein. Additionally, preisolation sera from each animal were able to neutralize the virus that caused the subsequent disease. These results provide evidence that antigenic variation is not a mechanism used by VS NJ virus to persist in an enzootic focus of Costa Rica.

Vesicular Stomatitis Virus, New Jersey Serotype: Replication in and Transmission by Lutzomyia shannoni (Diptera: Psychodidae)

Laboratory-reared female sand flies (Lutzomyia shannoni) were experimentally infected, orally and by intrathoracic inoculation, with the New Jersey serotype of vesicular stomatitis (VSNJ) virus. Virus replication occurred in the insects following infection by both routes. Virus titers greater than 10(4) plaque forming units of VSNJ virus were present in heads of orally infected sand flies 12 days after virus ingestion, confirming that a persistent disseminated infection had occurred. Both orally and parenterally infected Lu. shannoni transmitted VSNJ virus by bite to susceptible rodents and by transovarial transmission to a small percentage of their F1 progeny. The significance of these findings in the epizootiology of VSNJ virus on Ossabaw Island, Georgia, an enzootic focus of this virus, is discussed.

Caracterización cariológica de Proechimys sp. (Rodentia: Echimydae) de una colonia experimental

The Colombian National Institute of Health (INS) owns an experimental colony of Proechimys sp. which started from native animals from an enzootic focus located in Puerto Boyacá (Dos Quebradas area, Boyacá, Colombia). These animals have been used for viral replication of Venezuelan Equine Encephalitis virus since they are natural reservoirs of the infection. It this paper, we present the cariotype established by QFQ, GTC, CBG and NOR banding using lymphocyte microculture from peripheral blood. The chromosome number thus obtained (2n=32) is share with P. oconnelli (semispinosus group sensu Patton) having just about the same karyotype, and with P. simonsi (= P. hendeei, from the simonsi group sensu Patton) with remarkable differences When we analysed both the geograhical distribution and the karyology, we could think that even though they are very similar from the karyotypic point of view, P. oconnelli is probably not the species from this study The wide range of distribution of P. chrysaeolus could include the original location of this animals with wich the colony was started andy thereforey could be species that they belong to. However, its karyotype is unknown and its comparison become impossible The taxon of the Proechimys of the colony specifically will not be stablished unless the morphological and karyological knowledge is broadened on the species of the genus.

Comparison of Virulence between Two Strains of Rattus Serotype Hemorrhagic Fever with Renal Syndrome (HFRS) Virus in Newborn Rats

Two strains of hemorrhagic fever with renal syndrome (HFRS) virus from Rattus, SR-11 and KI-262, showed virtually identical antigenicity but differed from prototype strain Hantaan 76-118 (Apodemus origin) in a neutralization test. Wistar newborn rats inoculated intraperitoneally (i.p.) with SR-11, which was isolated from a laboratory rat associated with an outbreak of HFRS, developed clinical signs such as ataxia and limb paralysis and died at about 18 days after inoculation. The LD50 of SR-11 in 1-day-old rats was 10(1.2) focus-forming units (FFU). In contrast, the animals inoculated i.p. or intracerebrally with 10(4) FFU of KI-262, which was from a wild rat in a dumping-ground area--an enzootic focus where no human cases have been recorded--did not show any significant clinical signs. The susceptibility of rats to SR-11 fatal infection was age-dependent. Virus titers in brains, lungs, kidneys, and livers of the rats inoculated with SR-11 were significantly higher than those in the same organs of the animals infected with KI-262. Necrosis of neurons in the brain tissue occurred in the rats infected with SR-11, while it was mild in the animals infected with KI-262.

Infection status of Sesarma dehaani collected from Sumjin river delta with the metacercariae of Paragonimus iloktsuenensis

This study was performed to observe the recent infection status of Sesarma dehaani with the metacercariae of P. iloktsuenensis in the well-known enzootic focus, Sumjin river delta. A total of 74 Sesarma dehaani were collected from a focus near the mouth of the Sumjin river in November, 1986 and February, 1987. The crabs were examined for P. iloktsuenensis metacercariae by the method of Seo and Kwak(1972). The metacercariae of P. iloktsuenensis were found in the liver of the crabs. Among the 74 crabs examined, 47(63.5%) were found infected with 1-102 metacercariae (18.2 per crab). The infection rate and metacercarial density increased as the size of the crab was increased. From the results, it is suggested that the life cycle of P. iloktsuenensis is actively maintained in the Sumjin river basin.

Epizootiology of skunk rabies in North America.

Public health surveillance data from the United States and Canada (1961 to 1982) were analyzed to determine if consistent temporal and spatial patterns in skunk rabies could be identified. Enzootic/epizootic rabies was recognized in 18 states (enzootic states) based on the criteria of greater than or equal to 20 yr of reported skunk rabies and at least 1 yr with a minimum of 50 reported rabid skunks. In other wildlife species, epizootics have been demonstrated to expand along a wave-like front. We hypothesized: if skunk rabies behaved in a similar fashion, states reporting rabid skunks would change over time. No such change was noted. During epizootics the number of counties reporting increased but not the number of states. Within Illinois certain counties were demonstrated to have persistent rabies histories and likely served as enzootic foci. Enzootic states combined prevalence indicated a 6 to 8 yr cycle for epizootics. Data on monthly percent rabies positive (number rabid/total number tested) were available from six states and Canada. Mean distributions were bimodal with winter and spring peaks. The patterns identified for skunk rabies differ from those of other major wildlife vectors and have significance for potential vaccination control regimes.

Entomological studies at an enzootic Venezuelan equine encephalitis virus focus in Guatemala, 1977-1980.

The ecology of several potential mosquito vectors of Venezuelan equine encephalitis (VEE) alphavirus was studied in an enzootic focus of that virus on the Pacific coast of Guatemala over a four-year period. Four species-Culex taeniopus, Mansonia titillans, Culex nigripalpus and Aedes taeniorhynchus-were most prevalent during the wet season when transmission normally occurs. However, only Cx. taeniopus yielded VEE virus. The bloodfeeding patterns of these species revealed that Ae. taeniorhynchus and Ma. titillans fed almost exclusively on bovine and equine hosts. Conversely, Cx. nigripalpus was highly ornithophilic but occasionally fed on mammals. Cx. taeniopus exhibited a wide host range, utilizing both large and small mammals as well as birds and, rarely, reptiles. The versatility in feeding pattern displayed by this mosquito coupled with its ability to become infected with relatively low levels of enzootic VEE virus suggests that vertebrates other than rodents may serve as amplifying hosts in this habitat. Nepuyo virus was also isolated from Cx. taeniopus, suggesting that this mosquito might be an endemic vector of this rodent-associated bunyavirus. A single isolate of St. Louis encephalitis virus was made from Cx. nigripalpus.

Ecologic observations of Venezuelan encephalitis virus in vertebrates and isolations of Nepuyo and Patois viruses from sentinel hamsters at Pacific and Atlantic habitats in Guatemala, 1968-1980.

La Avellana and Puerto Barrios, two enzootic foci of Venezuelan encephalitis (VE) virus on the Pacific and Caribbean lowlands (respectively) of Guatemala have been studied over a 13-year period. Data from sentinel hamsters and guinea pigs and wild and domestic vertebrates are reported. VE virus strains were isolated from hamsters each period they were exposed during the rainy seasons 1968-1980 and at the end of the dry season 1974. Rates of isolation of VE virus ranged from 0.2%-5.7% hamster/days/exposure. All strains tested were free of epizootic virions. Although virus was isolated from sentinel guinea pigs, their deaths were not attributable to infection with VE virus. Antibody titers in 26 of 28 terrestrial mammals bled at La Avellana in 1971 were higher to enzootic than to epizootic VE strains. Thirty-seven percent of 109 residents of Puerto Barrios had antibody to VE virus. In 13 of 20 tested, antibodies were engendered by the enzootic strain. Nepuyo and Patois viruses were isolated from sentinel hamsters at both La Avellana and Puerto Barrios.

Arbovirus Studies in Southwestern Venezuela during 1973–1981

Increasing utilization of arable land in southwestern Venezuela has led to a potential increase in human exposure to arbovirus infections. Since previous studies in the Catatumbo region of this area documented the presence of eastern equine encephalitis (EEE) and Venezuelan equine encephalitis (VEE) viruses, an attempt was made to study the transmission and maintenance of these viruses from 1973 to 1981. Isolations of EEE, VEE ID strains, Una, Itaqui , and Moju viruses were repeatedly obtained from mosquitoes, mostly Culex ( Melanoconion ) spp. and sentinel hamsters. The results indicate that these viruses constitute a potential hazard to public health in the area. Further, the strategic location of the Catatumbo region, between enzootic tropical foci of arboviruses, may provide circumstances and conditions for study of both enzootic maintenance and movement of these viruses.

Blood-Feeding Patterns of Culiseta Melanura (Diptera: Culicidae) and Associated Sylvan Mosquitoes in Southeastern Massachusetts Eastern Equine Encephalitis Enzootic Foci1

The capillary precipitin technique was used to examine the host utilization pattern of Culiseta melanura in southeastern Massachusetts EEE enzootic foci. Resting boxes were used to collect 1792 blood-engorged females in sites at the perimeters of historically enzootic swamps, and 1563 in upland areas near horse corrals and human dwellings. Avian (primarily passerine) blood was identified in 98.7% of the meals at the perimeter during 1977, and in 98.8% outside the swamps in 1978. Nonavian single blood meals included: dog (0.22%); rabbit (0.3%); cat (0.6%); snake (0.06%); other reptiles (0.06%); and amphibians (0.03%). During 1978, 11.5% of the meals from a single perimeter site were mixed, frequently including rabbits and passerine birds. The general trend in both areas was toward almost exclusive feeding on passerines early in the season, with a gradual increase in feeding on other birds and, to a much lesser extent, mammals, reptiles, and amphibians later in the summer. Preliminary experiments showing greater fecundity with blood meals from chickens rather than rabbits indicate physiological as well as behavioral adaptations directed toward a successful relationship with avian hosts. Lack of feeding on EEE-susceptible mammals suggests that the role of Cs. melanura in the disease is limited to the avian cycle. Identifications of blood meals from other mosquito species point to Coquillettidia perturbans, Aedes canadensis, and Ae. vexans as possible vectors of EEE to horses and humans.

The epidemiology of tick-borne Crimean-Congo hemorrhagic fever in Asia, Europe, and Africa.

The epidemiology of tick-borne Crimean-Congo hemorrhagic fever in Asia, Europe, and Africa.

RODENT ZOONOSES IN NORTH QUEENSLAND

A study of potentially zoonotic infections was carried out on 351 rodents trapped in north-eastern Queensland. Their ecosystems included towns, agricultural and livestock areas, wookland and rainforest. Nine serotypes of salmonellae were obtained from asymptomatic carries in predominantly settled locations. Two strains of Ps. pseudomallei occurred in rainforest near Innisfail and one on a cattle property adjacent to Townsville. Ps. aeruginosa caused bronchopneumonia in one animal from Townsville harbour. Ifection by leptospirae of six serogroups and seven serovars were identified by serological or cultural examinations. Enzootic foci occurred on the Mount Spec rainforest where celledoni and australis were being excreted by rats adjacent to the Paluma dam system. In addition to the scrub typhus locations at Rocky Creek, Atherton Tableland and Bullocky Creek, near Ingham, which were confirmed, a new focus of infection by R. tsutsugamushi was identified at El Arish near Tully. Water rat (H. chrysogaster) at Townsville harbour constituted a reservoir of toxoplasmosis. In addition to the known human pathogenic helminths H. nana and H. diminuta, localized foci of hookworms (Ancylostoma spp.) were found. Histological evidence of cytomegalic disease of the salivary glands or kidneys was a common finding.

Epizootiology of epidemic typhus (Rickettsia prowazekii) in flying squirrels.

Vector transmission of Rickettsia prowazekii among wild flying squirrels, Glaucomys volans, was suggested by the occurrence of natural infection of squirrel lice and fleas. Lice, mostly Neohaematopinus sciuropteri Osburn, were found infected in the fall in each of 2 consecutive years; 4 of the 8 pools of this insect tested were infected. Fleas, Orchopeas howardii (Baker), were found infected on two occasions in 1 of the 2 consecutive years. However, only 2 of 14 flea pools were infected. No evidence of infection was found in mites, Haemogamasus reidi Ewing and Androlaelaps fahrenholzi (Berlese). These findings implicate the flying squirrel louse and flea as possible vectors in nature. Serologic tests of flying squirrel sera revealed a maximum incidence of seroconversions in the fall and early winter months, coincident with the maximum increase in abundance of the suspected arthropod vectors. The infection was found to persist form year to year in the same enzootic foci. Infection appeared to spread most rapidly in young, non-immune animals born in the preceding spring and summer after congregating in dense aggregations in the fall. No other animals in the same habitat were found to have been infected. Aspects of the ecology of the ectoparasites associated with the flying squirrels are described, especially seasonal activity and abundance in nests. The potential public health importance of this sylvan disease in flying squirrels and in its ectoparasites, particularly the non-host specific, wide ranging squirrel flea, is noted.

Venezuelan encephalitis virus infection in neotropical bats. I. Natural infection in a Guatemalan enzootic focus.

A serological survey of 939 Neotropical bats of 22 species from an enzootic focus of Venezuelan encephalitis (VE) virus on the Pacific lowlands of Guatemala during 1971--1975 revealed VE virus specific antibodies in seven species, three belonging to the genus Artibeus. VE virus was isolated from the blood of one Uroderma bilobatum. Antibody frequency was considerably lower in bats than in terrestrial mammals, and tended to vary within any given species from locality to locality and from year to year. At the village of La Avellana where VE virus was most active, antibody rates in the two best-sampled Artibeus species were 0.14 in 1971, 0.11 in 1972, 0.03 in 1973, and 0.11 in 1975. The positive rate of 0.10 over all years at La Avellana for the genus Artibeus suggests that these and possibly other bats regularly are infected by VE virus, and may possibly serve as alternate hosts to maintain virus circulation if most terrestrial animals become immune.

Search for subtype I strains of Venezuelan encephalitis virus among isolates from Trinidad, W.I., during 1960-1972.

Twenty-one strains of Venezuelan encephalitis (VE) virus isolated from three habitats in Trinidad, W.I. during 1960 to 1972, were subtype III (Mucambo) VE virus by plaque-reduction neutralization tests. Like prototype Mucambo virus, each strain killed 8- to 15-week-old mice inoculated intraperitoneally. If the subtype I strain of VE virus that caused a major outbreak in Trinidad during 1943 to 1944 persisted on the island into the 1960s and early 1970s, it did not become the dominant VE virus in these three enzootic foci.

Human Babesiosis on Nantucket Island: Transmission by Nymphal Ixodes Ticks

In order to identify potential vectors of human babesiosis (Babesia microli) in southern Massachusetts, I attempted to transmit the infection via nymphal ticks of that species found to be most abundant on reservoir hosts (Peromyscus leucopus) in nature. Mice were collected at frequent intervals throughout a year in a known enzootic focus on Nantucket Island, and ticks were removed by hand. Ticks of only two species were present; larvae and nymphs of Ixodes scapularis were about 10 times as numerous as were those of Dermacentor variabilis. Accordingly, I. scapularis were used in attempts to transmit a strain of Babesia derived from a human infection recently acquired near the study site. Larvae were permitted to feed on an infected hamster and nymphs derived from these larvae were placed on each of 11 non-infected hamsters. All but one hamster became infected. Nymphs reared from larvae that had attached to a non-infected hamster did not transmit babesiosis to other hamsters. These results suggest that I. scapularis serves as a vector of babesios-s on Nantucket hamsters. These results suggest that I. scapularis serves as a vector of babesiosis on Nantucket Island.

Search for epizootic-like Venezuelan encephalitis virus at enzootic habitats in Guatemala during 1969-1971.

Seventy-four strains of Venezuelan encephalitis (VE) virus recovered from sentinel hamsters or mosquitoes at enzootic habitats in Guatemala in the two years following the 1969 epidemic-equine epizootic were examined for ability to produce small plaques in Vero African green monkey kidney cell cultures, like isolates obtained during the epizootic. (a) One strain recovered from a sentinel hamster in late October 1969 at an enzootic habitat near the epicenter of the hemagglutination-inhibition (HI) and equine-virulence properties like epizootic virus; this strain retained its small plaque characteristic after inoculation and recovery from bloods of three horses. (b) None of the other 73 strains produced uniformly small plaques, but 31 formed a few small plaques among large ones. Virions from small plaques of five strains were cloned twice in Vero cell cultures. Four clones produced uniformly small plaques after one more passage in Vero cells; three had hemagglutination-pH properties compatible with epizootic virus or intermediate between epizootic and enzootic virus, but HI tests with these three hemagglutinins or with antibody to the fourth cloned strain showed them to be like Central American enzootic virus. One of three cloned strains tested in horses produced encephalitis and death in one of four horses; another strain produced encephalitis with recovery in one of two horses. (c) Thus these small Vero plaque clones resembled Central American enzootic strains of VE virus in HI and equine-virulence tests, and the small Vero plaque characteristic was not a satisfactory marker for consistently isolating equine-virulent, epizootic VE virions. Nevertheless, this technic led to recognition of one epizootic strain isolated at an enzootic habitat in Guatemala at the end of 1969 outbreak. Whether this strain was there before the outbreak or subsequently penetrated the habitat is uncertain. During the next two years, this strain did not become dominant in that enzootic focus.

Studies of possible movement of Venezuelan encephalitis virus from an enzootic focus in Guatemala during 1971-1974.

During the wet seasons of 1972 and possibly 1971, sentinel horses became infected by Venezuelan encephalitis (VE) virus in a temporally and geographically progressive manner inland from an enzootic marsh focus of virus on the Pacific couast of southeastern Guatemala. During the wet seasons of 1972 and 1973, VE virus was detected by sentinel horses (and a sentinel hamster in 1972) in a small woods 10 km north of the marsh, but virus was undetectable there during the dry seasons of 1973 and 1974 and the wet season of 1974. Culex (Melanoconion) mosquitoes were found in this woods and at the marsh during August 1973. These observations are compatible with movement of VE virus from the marsh habitat during some wet seasons. However, virus activity in this region adjacent to the marsh was quantitatively unpredictable on a yearly basis and occurred in only very focal habitats during 1971 to 1974. Mechanisms of VE virus movement from the marsh are currently unknown, but bats are under study as a likely possibility.