Role In Transmission Of Virus Research Articles

Practical Mitigation Strategies for Countering the Spread of Aerosolized COVID-19 Virus (SARS-CoV-2) Using Ventilation and HEPA Air Purifiers: A Literature Review

This paper assesses, through an extensive literature review, the use of ventilation and High-Efficiency Particulate Air (HEPA) purifiers as practical mitigation strategies for reducing the spread of aerosolized COVID-191 virus. HEPA is a well-defined standard by the U.S. Department of Energy for filters. The focus of the literature review was on indoor air quality (IAQ) and COVID-19, with a particular emphasis on classroom settings. The start of the review, January 2020, was chosen to coincide with the first cases of COVID-19 in North America. Although children under the age of 12 are currently not yet vaccinated, there is mounting pressure for a return to normal by the start of the new school year, 2021. Also, many classrooms lack pre-installed mechanical ventilation systems (Olsiewski et al., 2021); therefore, mitigation in classrooms often falls solely in the hands of teachers and students. Research shows that ventilation and air purification are essential tools to counter aerosolized transmission (Curtius et al. (2020), the inhaled dose of particles containing virus RNA is six times lower when using air purifiers with an ACH (air changes per hour) of 5.7. However, ventilation and air purifiers are not replacements for masks, which remain vital for countering droplet (>5 μm) transmission. In addition, occupancy (i.e., number and proximity of people present in a given area) and group activity levels (e.g., talking, shouting, singing) play a critical role in viral transmission. Although natural ventilation by opening windows can be an essential strategy to help counter the spread of the virus, the level of ventilation offered by opening windows is largely uncontrollable as it is subject to weather conditions and building design. One must also consider the energy implications (i.e., loss of heat) that this strategy carries. Scientific evidence shows that varying levels of continuous and/or intermittent ventilation, either mechanical or natural, combined with the use of HEPA air purifiers, can provide a higher degree of protection than window access alone (Curtius et al., 2020). Systematic deployment of a hybrid mitigation strategy incorporating both ventilation and HEPA air purification in schools, offices, or other facilities offers a practical way to establish a safe re-opening of society in Canada.

Children experienced new or worsening tic issues when they were separated from their parents during the Italian COVID-19 lockdown.

Decades of research have confirmed that children develop best when they are in safe, supportive, nurturing relationships (1). Positive relationships with parents are essential for children's growth, health and wellbeing (1,2), as they protect them and help them to regulate their emotions during stressful times. If children are unnecessarily and traumatically removed from their parents, their physical and mental health and wellbeing will suffer (1,2).

Clinical characteristics of asymptomatic carriers of novel coronavirus disease 2019: A multi-center study in Jiangsu Province

ABSTRACT Asymptomatic SARS-CoV-2-infected individuals are thought to play major roles in virus transmission. This study aimed to analyze the characteristics of asymptomatic carriers with COVID-19 to control the spread of the virus. We retrospectively investigated the clinical characteristics of 648 consecutive subjects who were enrolled in the study and were divided into asymptomatic carriers, mild cases, ordinary cases, severe or critical cases, and evaluated their impact on disease severity by means of Spearman correlation and multiple regression analyses. Receiver operating characteristic curve analysis was conducted to determine the optimum cutoff levels of laboratory findings for diagnostic predictors of asymptomatic carriers of COVID-19. In our study, a total of 648 subjects on admission with a mean age of 45.61 y including 345 males and 303 females were enrolled in our study. The leukocyte, lymphocyte, eosinophil, platelet, C-reactive protein, interleukin-6, CD3+, CD4+, and CD8 + T lymphocyte levels, and the erythrocyte sedimentation rate differed significantly among the groups (all p ≤ 0.05). Disease severity was negatively associated with the CD3+ (r = −0.340; p < 0.001), CD4+ (r = −0.290; p = 0.001) and CD8+ (r = −0.322; p < 0.001) T lymphocyte levels. The significant diagnostic predictors of asymptomatic carriers of COVID-19 included the blood cell, cytokine, and T lymphocyte subset levels. Inflammation and immune response may play important roles in disease progression. Hence, the laboratory parameters identified should be considered in clinical practice, which provide new insights into the identification of asymptomatic individuals and the prevention of virus transmission.

Characterization of flavivirus infection in salivary gland cultures from male Ixodes scapularis ticks.

Infected Ixodes scapularis (black-legged tick) transmit a host of serious pathogens via their bites, including Borrelia burgdorferi, Babesia microti, and tick-borne flaviviruses (TBFVs), such as Powassan virus (POWV). Although the role of female I. scapularis ticks in disease transmission is well characterized, the role of male ticks is poorly understood. Because the pathogens are delivered in tick saliva, we studied the capacity of male salivary glands (SGs) to support virus replication. Ex vivo cultures of SGs from unfed male I. scapularis were viable for more than a week and maintained the characteristic tissue architecture of lobular ducts and acini. When SG cultures were infected with the TBFVs Langat virus (LGTV) or POWV lineage II (deer tick virus), the production of infectious virus was demonstrated. Using a green fluorescent protein-tagged LGTV and confocal microscopy, we demonstrated LGTV infection within SG acinus types II and III. The presence of LGTV in the acini and lobular ducts of the cultures was also shown via immunohistochemistry. Furthermore, the identification by in situ hybridization of both positive and negative strand LGTV RNA confirmed that the virus was indeed replicating. Finally, transmission electron microscopy of infected SGs revealed virus particles packaged in vesicles or vacuoles adjacent to acinar lumina. These studies support the concept that SGs of male I. scapularis ticks support replication of TBFVs and may play a role in virus transmission, and further refine a useful model system for developing countermeasures against this important group of pathogens.

Coronaviruses and Endocrine System: A Systematic Review on Evidence and Shadows.

Coronaviruses are a big family of viruses that can infect mammalians and birds. In humans they mainly cause respiratory tract infections, with a large spectrum of severity, from mild, self-limited infections to highly lethal forms as severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV) and Coronavirus Disease 2019 (COVID-19). Scanty data are reported for the involvement of endocrine glands in human coronaviruses, in particular SARS-CoV-2. In this review, we summarize endocrinological involvement in human coronaviruses, including data on animal coronaviruses. Avians, ferrets and bovine are affected by specific coronavirus syndromes, with variable involvement of endocrine glands. SARS-CoV and SARS-CoV-2 use angiotensin-converting enzyme 2 (ACE2) as a target receptor, so ACE2 plays a central role in viral transmission and initial organ involvement. Autoptic studies on SARS patients revealed that thyroid, parathyroid, pituitary gland, endocrine pancreas and especially adrenals and testis could be impaired by different mechanisms (direct damage by SARS-CoV, inflammation, vascular derangement and autoimmune reactions) and few clinical studies have evidenced functional endocrine impairment. Only few data are available for COVID-19 and gonads and endocrine pancreas seem to be involved. International endocrinological societies have brought some recommendations for the COVID-19 pandemic, but further studies need to be performed, especially to detect long-term hormonal sequelae.

Human immunodeficiency virus interaction with oral and genital mucosal epithelia may lead to epithelial-mesenchymal transition and sequestration of virions in the endosomal compartments.

Oral and genital mucosal epithelia are multistratified epithelial barriers with well-developed tight and adherens junctions. These barriers serve as the first line of defense against many pathogens, including human immunodeficiency virus (HIV). HIV interaction with the surface of mucosal epithelial cells, however, may activate transforming growth factor-beta (TGF-β) and mitogen-activated protein kinase signaling pathways. When activated, these pathways may lead to the disruption of epithelial junctions and epithelial-mesenchymal transition (EMT). HIV-induced impairment of the mucosal barrier may facilitate the spread of pathogenic viral, bacterial, fungal, and other infectious agents. HIV-induced EMT promotes highly motile/migratory cells. In oral and genital mucosa, if EMT occurs within a human papillomavirus (HPV)-infected premalignant or malignant cell environment, the HPV-associated neoplastic process could be accelerated by promoting viral invasion of malignant cells. HIV also internalizes into oral and genital mucosal epithelial cells. The majority (90%) of internalized virions do not cross the epithelium, but are retained in endosomal compartments for several days. These sequestered virions are infectious. Upon interaction with activated peripheral blood mononuclear cells and CD4+ T lymphocytes, epithelial cells containing the virus can be transferred. The induction of HIV-1 release and the cell-to-cell spread of virus from epithelial cells to lymphocytes is mediated by interaction of lymphocyte receptor function-associated antigen-1 with the epithelial cell receptor intercellular adhesion molecule-1. Thus, mucosal epithelial cells may serve as a transient reservoir for HIV, which could play a critical role in viral transmission.

COVID-19 Pandemic and Role of Human Saliva as a Testing Biofluid in Point-of-Care Technology.

Novel coronavirus disease 2019 (COVID-19) outbreak has termed as a controllable pandemic, and the entire world has come to a standstill trying to mitigate the disease with health systems. Health care providers, around the globe, are fighting day and night. Currently, rapid testing is taking place with the help of nasopharyngeal, oropharyngeal swab, bronchoalveolar lavage, sputum, urine, and blood. All these approaches are invasive or embarrassing to the infected person. It is observed that salivary glands are hosting severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) because of angiotensin-converting enzyme 2 and the detection of high viral loads in the saliva and is playing a crucial role in virus transmission, especially from individuals showing absolutely no symptoms. Saliva is proving to be a promising noninvasive sample specimen for the diagnosis of COVID-19, thus helping to monitor the infection and prevent it from further spreading by prompt isolation.

Innate and Adaptive Anti-SIV Responses in Macaque Semen: Implications for Infectivity and Risk of Transmission.

HIV-1 infection is transmitted primarily by sexual exposure, with semen being the principal contaminated fluid. However, HIV-specific immune response in semen has been understudied. We investigated specific parameters of the innate, cellular, and humoral immune response that may affect semen infectivity in macaques infected with SIVmac251. Serial semen levels of cytokines and chemokines, SIV-specific antibodies, neutralization, and FcγR-mediated functions and SIV-specific T-cell responses were assessed and compared to systemic responses across 53 cynomolgus macaques. SIV infection induced an overall inflammatory state in the semen. Several pro-inflammatory molecules correlated with SIV virus levels. Effector CD8+ T cells were expanded in semen upon infection. SIV-specific CD8+ T-cells that expressed multiple effector molecules (IFN-γ+MIP-1β+TNF+/−) were induced in the semen of a subset of SIV-infected macaques, but this did not correlate with local viral control. SIV-specific IgG, commonly capable of engaging the FcγRIIIa receptor, was detected in most semen samples although this positively correlated with seminal viral load. Several inflammatory immune responses in semen develop in the context of higher levels of SIV seminal plasma viremia. These inflammatory immune responses could play a role in viral transmission and should be considered in the development of preventive and prophylactic vaccines.

Historical origins and zoonotic potential of avian influenza virus H9N2 in Tunisia revealed by Bayesian analysis and molecular characterization.

During 2009-2012, several outbreaks of avian influenza virus H9N2 were reported in Tunisian poultry. The circulating strains carried in their hemagglutinins the human-like marker 226L, which is known to be important for avian-to-human viral transmission. To investigate the origins and zoonotic potential of the Tunisian H9N2 viruses, five new isolates were identified during 2012-2016 and their whole genomes were sequenced. Bayesian-based phylogeny showed that the HA, NA, M and NP segments belong to the G1-like lineage. The PB1, PB2, PA and NS segments appeared to have undergone multiple intersubtype reassortments and to be only distantly related to all of the Eurasian lineages (G1-like, Y280-like and Korean-like). The spatiotemporal dynamic of virus spread revealed that the H9N2 virus was transferred to Tunisia from the UAE through Asian and European pathways. As indicated by Bayesian analysis of host traits, ducks and terrestrial birds played an important role in virus transmission to Tunisia. The subtype phylodynamics showed that the history of the PB1 and PB2 segments was marked by intersubtype reassortments with H4N6, H10N4 and H2N2 subtypes. Most of these transitions between locations, hosts and subtypes were statistically supported (BF > 3) and not influenced by sampling bias. Evidence of genetic evolution was observed in the predicted amino acid sequences of the viral proteins of recent Tunisian H9N2 viruses, which were characterized by the acquisition of new mutations involved in virus adaptation to avian and mammalian hosts and amantadine resistance. This study is the first comprehensive analysis of the evolutionary history of Tunisian H9N2 viruses and highlights the zoonotic risk associated with their circulation in poultry, indicating the need for continuous surveillance of their molecular evolution.

Deposition of bronchiole-originated droplets in the lower airways during exhalation

Recent studies have shown that small expired droplets that contain pathogens may play a significant role in the transmission of respiratory diseases. However, there is little data on the generation of droplets from the bronchioles and their deposition in the lower airways during exhalation. We apply a computational fluid dynamics method to study expiratory air flow in a truncated model of the adult lower airways. Assumptions are made to evaluate the transport and deposition of droplets along the airway as these droplets are released in various regions in the conducting airways during exhalation. We find that large droplets (i.e., >5 μm) generated from the bronchioles are very likely to be blocked by the lower airways, whereas those in the submicron range (i.e., <1 μm) easily survive the conducting tracts and are likely to be exhaled. The locations at which large droplets from the bronchioles are deposited are limited to the terminal regions, which means that these droplets are unlikely to be released again via possible aerosolization due to the shear force on the airway walls. The evidence from the computational fluid dynamics simulation suggests that particle size plays an important role in virus transmission and disease diagnosis.

Climate variability and seasonal weather related to COVID-19

In view of the alarming situation related to coronavirus, of particular interest to the public, decision makers, health organizations, experts, professionals (epidemiologists, virologists, infectologists, psychologists, pulmonologists and others), it is also important to obtain seasonal weather outlook with general overview of the weather and climate conditions and how they would potentially affect the state of the COVID-19 transmission because a number of infectious diseases show seasonal features in their incidence. The analysis below represents an attempt to evaluate atmospheric processes that have a different spatiotemporal scale and depend on other modes of climate variability, and patterns of circulation and specific indices that affect seasonal variation and weather characteristics in a region. Based on seasonal outlook, the general impression is that sunny, warmer increase of (1.0 to 2.0°C) and frequently stormy weather conditions expected during May-June 2020 could likely give some optimistic scenario of slowing down the spreading of the virus further. This is confirmed with the further evaluation of the effective reproduction number, that is expected to decrease from 1.8 at middle of April to 1.2 at the end of June 2020. That is approximately 60-70 % reduction during this warmer period. However, the conclusions are still general and should be taken with caution as the situation is changing from day to day and many other factors including climate conditions, population density and medical care quality also play role in virus transmission.

Influenza in Children.

Influenza poses a significant disease burden on children worldwide, with high rates of hospitalization and substantial morbidity and mortality. Although the clinical presentation of influenza in children has similarities to that seen in adults, there are unique aspects to how children present with infection that are important to recognize. In addition, children play a significant role in viral transmission within communities. Growing evidence supports the idea that early influenza infection can uniquely establish lasting immunologic memory, making an understanding of how viral immunity develops in this population critical to better protect children from infection and to facilitate efforts to develop a more universally protective influenza vaccine.

From a Movement-Deficient Grapevine Fanleaf Virus to the Identification of a New Viral Determinant of Nematode Transmission.

Grapevine fanleaf virus (GFLV) and arabis mosaic virus (ArMV) are nepoviruses responsible for grapevine degeneration. They are specifically transmitted from grapevine to grapevine by two distinct ectoparasitic dagger nematodes of the genus Xiphinema. GFLV and ArMV move from cell to cell as virions through tubules formed into plasmodesmata by the self-assembly of the viral movement protein. Five surface-exposed regions in the coat protein called R1 to R5, which differ between the two viruses, were previously defined and exchanged to test their involvement in virus transmission, leading to the identification of region R2 as a transmission determinant. Region R4 (amino acids 258 to 264) could not be tested in transmission due to its requirement for plant systemic infection. Here, we present a fine-tuning mutagenesis of the GFLV coat protein in and around region R4 that restored the virus movement and allowed its evaluation in transmission. We show that residues T258, M260, D261, and R301 play a crucial role in virus transmission, thus representing a new viral determinant of nematode transmission.

Quantity and transmission efficiency of an isolate of the Potato virus Y–Wilga (PVYN−Wi) by aphid species reared on different host plants

Potato virus Y (PVY) is a destructive plant virus causing important damage in different crops, particularly in potato. PVY is transmitted in a non-persistent manner by many aphid species, and the distal part of the stylet plays an important role in virus transmission by these aphids. Among other possible factors, the type of host plant on which the aphids develop could also affect virus transmission efficiency, yet little is known about this. Furthermore, the quantification of potyviruses in individual vectors has mainly focused on intact whole aphid body and not the stylet alone, whereas an even more precise indicator that could correlate with vector efficiency in non-persistently transmitted viruses is the quantity of the virus in the stylet. Therefore, this study aims to verify the effect of different host plants on which an aphid vector was reared on its ability to acquire and transmit the PVY, and whether the quantity of PVY in the aphid vector stylet could be used as a useful trait for evaluating vector efficiency. A key finding in this study indicates that when the aphid Myzus persicae was reared on broad bean and then exposed to acquire PVY, it transmitted PVY less efficiently (50%) compared to when it was reared on Brussels sprout (81%). Additionally, the transmission percentage correlated directly with the virus quantity in the stylet (r = 0.7) and not in the intact whole aphid body (r = 0.06). This could be exploited as a useful index for virus transmission efficiency in epidemiological studies or predictive schemes.

Exploring the Reservoir Hosts of Tick-Borne Encephalitis Virus.

Tick-borne encephalitis virus (TBEV) is an important arbovirus, which is found across large parts of Eurasia and is considered to be a major health risk for humans. Like any other arbovirus, TBEV relies on complex interactions between vectors, reservoir hosts, and the environment for successful virus circulation. Hard ticks are the vectors for TBEV, transmitting the virus to a variety of animals. The importance of these animals in the lifecycle of TBEV is still up for debate. Large woodland animals seem to have a positive influence on virus circulation by providing a food source for adult ticks; birds are suspected to play a role in virus distribution. Bank voles and yellow-necked mice are often referred to as classical virus reservoirs, but this statement lacks strong evidence supporting their highlighted role. Other small mammals (e.g., insectivores) may also play a crucial role in virus transmission, not to mention the absence of any suspected reservoir host for non-European endemic regions. Theories highlighting the importance of the co-feeding transmission route go as far as naming ticks themselves as the true reservoir for TBEV, and mammalian hosts as a mere bridge for transmission. A deeper insight into the virus reservoir could lead to a better understanding of the development of endemic regions. The spatial distribution of TBEV is constricted to certain areas, forming natural foci that can be restricted to sizes of merely 500 square meters. The limiting factors for their occurrence are largely unknown, but a possible influence of reservoir hosts on the distribution pattern of TBE is discussed. This review aims to give an overview of the multiple factors influencing the TBEV transmission cycle, focusing on the role of virus reservoirs, and highlights the questions that are waiting to be further explored.

Evolution-Driven Attenuation of Alphaviruses Highlights Key Glycoprotein Determinants Regulating Viral Infectivity and Dissemination

Understanding the fundamental mechanisms of arbovirus transmission and pathogenesis is essential to develop strategies for treatment and prevention. We previously took an invivo evolution-based approach and identified the chikungunya virus E1 glycoprotein residue 80 to play a critical role in viral transmission and pathogenesis. In this study, we address the genetic conservation and function of position 80 and demonstrate that this residue is a key determinant in alphavirus infectivity and dissemination through modulation of viral fusion and cholesterol dependence. In addition, in studying the evolution of position 80, we identified a network of glycoprotein residues, including epidemic determinants, that regulate virus dissemination and infectivity. These studies underscore the importance of taking evolution-based approaches to not only identify key viral determinants driving arbovirus transmission and pathogenesis but also to uncover fundamental aspects of arbovirus biology.

Plant-Mediated Silencing of the Whitefly Bemisia tabaci Cyclophilin B and Heat Shock Protein 70 Impairs Insect Development and Virus Transmission.

The whitefly B. tabaci is a global pest and transmits extremely important plant viruses especially begomoviruses, that cause substantial crop losses. B. tabaci is one of the top invasive species worldwide and have developed resistance to all major pesticide classes. One of the promising alternative ways for controlling this pest is studying its genetic makeup for identifying specific target proteins which are critical for its development and ability to transmit viruses. Tomato yellow leaf curl virus (TYLCV) is the most economically important and well-studied begomovirus transmitted by B. tabaci, in a persistent-circulative manner. Recently, we reported that B. tabaci Cyclophilin B (CypB) and heat shock protein 70 proteins (hsp70) interact and co-localize with TYLCV in the whitefly midgut, on the virus transmission pathway, and that both proteins have a significant role in virus transmission. Here, we extended the previous work and used the Tobacco rattle virus (TRV) plant-mediated RNA silencing system for knocking down both genes and testing the effect of their silencing on whitefly viability and virus transmission. Portions of these two genes were cloned into TRV constructs and tomato plants were infected and used for whitefly feeding and transmission experiments. Following whitefly feeding on TRV-plants, the expression levels of cypB and hsp70 in adult B. tabaci significantly decreased over 72 h feeding period. The knockdown in the expression of both genes was further shown in the first generation of silenced whiteflies, where phenotypic abnormalities in the adult, wing, nymph and bacteriosomes development and structure were observed. Additionally, high mortality rates that reached more than 80% among nymphs and adults were obtained. Finally, silenced whitefly adults with both genes showed decreased ability to transmit TYLCV under lab conditions. Our results suggest that plant-mediated silencing of both cypB and hsp70 have profound effects on whitefly development and its ability to transmit TYLCV.

How Host Specific Are Herpesviruses? Lessons from Herpesviruses Infecting Wild and Endangered Mammals.

Herpesviruses are ubiquitous and can cause disease in all classes of vertebrates but also in animals of lower taxa, including molluscs. It is generally accepted that herpesviruses are primarily species specific, although a species can be infected by different herpesviruses. Species specificity is thought to result from host-virus coevolutionary processes over the long term. Even with this general concept in mind, investigators have recognized interspecies transmission of several members of the Herpesviridae family, often with fatal outcomes in non-definitive hosts-that is, animals that have no or only a limited role in virus transmission. We here summarize herpesvirus infections in wild mammals that in many cases are endangered, in both natural and captive settings. Some infections result from herpesviruses that are endemic in the species that is primarily affected, and some result from herpesviruses that cause fatal disease after infection of non-definitive hosts. We discuss the challenges of such infections in several endangered species in the absence of efficient immunization or therapeutic options.

Mouse mammary tumour virus-like env nucleotide and p14 signal peptide are present in feline mammary carcinomas, but not in neoplastic or dysplastic canine mammary lesions.

Mouse mammary tumour virus-like (MMTV-like) is suspected to be involved in human breast cancer and it has been hypothesized that companion animals might have a role in viral transmission. The aim of our study was to investigate the presence of MMTV-like nucleotide sequences and viral protein in a larger number of feline (FMCs) and canine mammary carcinomas (CMCs) by nested PCR and immunohistochemistry. Results showed that the presence of MMTV-like env sequence in FMCs was 7% (6/86), while all the CMCs and canine dysplastic lesions scored negative. All PCR-positive FMCs scored positive for the MMTV p14 signal peptide of the envelope precursor protein of the virus. In contrast, all PCR-negative FMCs and canine mammary lesions were also negative for immunohistochemistry analysis. Canine and feline normal mammary gland tissues scored negative for both PCR and MMTV-p14 protein. Multiple nucleotide alignment of MMTV-like env gene sequences isolated from cat showed 97% and 99% similarity with HMTV and MMTV, respectively, while the others two presented some polimorphisms. Particularly the sequences of one of these two tumors showed a polymorphism (c.7575 A> G), that causes a previously unreported amino acid substitution (Thr > Ala). In conclusion, the results of our study showed the presence of MMTV-like sequences and viral protein in some FMCs. Further studies are needed to understand whether this virus does play a role in the development of FMCs, if MMTV-like is an exogenous virus as these data suggest and, in such a case, how and from whom this virus was acquired.

Seasonal Distribution and Container Preference Ratio of the Dengue Fever Vector (Aedes aegypti, Diptera: Culicidae) in Rawalpindi, Pakistan.

Aedes aegypti (L.) and Aedes albopictus (Skuse) are known vectors of dengue, chikungunya, and other pathogens; however, their ecology and role in virus transmission has not been well studied in Pakistan. Here, we report on an intensive survey of potential breeding sites of Ae. aegypti in Rawalpindi, Punjab Province, Pakistan. The study continued for 11 mo and was divided into three seasons: January to June (pre-monsoon), July to September (monsoon), and October-November (post-monsoon). Larval mosquitoes were collected from all wet containers present in and around the houses. Altogether 5,570,418, 2,930,508, and 1,507,111 water-filled containers were examined during each season, of which 2,703, 8,843, and 3,439 were found positive for Ae. aegypti larvae or pupae, yielding Breteau indices of 0.46, 2.92, and 1.99%, respectively. Among 14 container types examined, the breeding preference ratio during all seasons was highest for roof-top water tanks and room evaporative coolers, followed by discarded tires and urban trash. The study concluded that increased urbanization, insufficient water supply and inefficient removal of urban trash resulted in increased numbers of nonbiodegradable containers around human dwellings, thereby creating ideal breeding habitats for Ae. aegypti. Measures such as integrated vector management, minimization of the breeding potential of Ae. aegypti by water management, proper disposal of discarded tires and urban trash, and health education were recommended for control of Ae. aegypti.