Scent, sweat from human skin attract disease-spreading mosquitoes

Mosquitoes that spread Zika, dengue and yellow fever are guided toward their victims by a scent from human skin. The exact composition of that scent has not been identified until now.

A UC Riverside-led team discovered that the combination of carbon dioxide plus two chemicals, 2-ketoglutaric and lactic acids, elicits a scent that causes a mosquito to locate and land on its victim. This chemical cocktail also encourages probing, the use of piercing mouthparts to find blood.

This chemical mixture appears to specifically attract female Aedes aegypti mosquitoes, vectors of Zika as well as chikungunya, dengue, and yellow fever viruses. This mosquito originated in Africa, but has spread to tropical and subtropical regions worldwide, including the U.S.

Mosquitoes use a variety of cues to locate their victims, including carbon dioxide, sight, temperature, and humidity. However, Cardé’s recent research shows skin odors are even more important for pinpointing a biting site.

Mosquito

Aedes aegyptii mosquito biting a person./CREDIT CDC

“We demonstrated that mosquitoes land on visually indistinct targets imbued with these two odors, and these targets aren’t associated with heat or moisture,” Cardé said. “That leaves skin odor as the key guiding factor.”

Given the significance of odor in helping mosquitoes successfully feed on humans, Cardé wanted to discover the exact chemicals that make our scent so potent for the insects. Part of the equation, lactic acid, was identified as one chemical element in the odor cocktail as long ago as 1968.

Since then, several studies have identified that carbon dioxide combined with ammonia, and other chemicals produced by humans also attract these mosquitoes. However, Cardé, who has studied mosquitoes for 26 years, felt these other chemicals were not strong attractants.

Methods that chemists typically use to identify these chemicals would not have worked for 2-ketoglutaric acid, Cardé said. Gas chromatography, which separates chemicals by their molecular weight and polarity, would have missed this acid.

“I think that these chemicals may not have been found before because of the complexity of the human odor profile and the minute amounts of these compounds present in sweat,” said chemist Jan Bello, formerly of UCR and now with insect pest control company Provivi.

Searching for mosquito attractors, Cardé turned to Bello, who extracted compounds from the sweat in his own feet. He filled his socks with glass beads and walked around with the beads in his socks for four hours per odor collection.

Malaria

Credit: MINDY TAKAMIYA/KYOTO UNIVERSITY ICEMS

“Wearing the beads felt almost like a massage, like squeezing stress balls full of sand, but with your feet,” said Bello. ‘The most frustrating part of doing it for a long time is that they would get stuck in between your toes, so it would be uncomfortable after a while.”

The inconvenience was worth the investment. Bello isolated chemicals from the sweat deposited on the sock beads and observed the mosquitoes’ response to those chemicals. In this way, the most active combination emerged.

Future studies are planned to determine whether the same compound is effective for any other mosquitoes, and why there is such variation in how individuals are apt to be bitten. “Some are more attractive than others to these mosquitoes, but no one’s yet established why this is so,” Cardé said.

Though this discovery may not lead to insights for the development of new repellants, the research team is hopeful their discovery can be used to attract, trap, and potentially kill disease-spreading mosquitoes.

Making mosquitoes self-destruct

Researchers at the University of California, Riverside have developed transgenic mosquitoes that stably express the Cas9 enzyme in their germline. The addition of Cas9 will enable the use of the CRISPR gene editing tool to make efficient, targeted changes to the mosquitoes’ DNA.

As proof of concept, the researchers used the system to disrupt cuticle, wing, and eye development, producing completely yellow, three-eyed and wingless mosquitoes. Their long-term goal is to use Cas9-expressing mosquitoes together with another technology — called gene drives — to insert and spread genes that suppress the insects while avoiding the resistance that evolution would typically favor. Aedes aegypti are major carriers of dengue, chikungunya, yellow fever, and zika viruses, and are rapidly becoming resistant to commonly used pesticides.

Published today in the Proceedings of the National Academy of Sciences (PNAS), the study was led by Omar Akbari, an assistant professor of entomology in UCR’s College of Natural and Agricultural Sciences and a member of the university’s Institute for Integrative Genome Biology.

Previous efforts to use genome editing to prevent mosquitoes from spreading pathogens have been hampered by low mutation rates, poor survival of edited mosquitoes, and inefficient transmission of disrupted genes to offspring. Akbari and colleagues developed transgenic mosquitoes that stably express a bacterial Cas9 enzyme in the germline, enabling highly efficient genome editing using the CRISPR system. CRISPR works like a pair of molecular scissors, cutting out and replacing specific DNA sequences based on a ribonucleic acid (RNA) guide. In the paper, the team used the system to disrupt genes that control vision, flight and feeding, resulting in mosquitoes with an extra eye, malformed wings, and defects in eye and cuticle color, among other changes.

Akbari said these strains represent the first step toward using gene drive systems to control mosquito populations and reduce the diseases they spread.

“These Cas9 strains can be used to develop split-gene drives which are a form of gene-drive by which the Cas9 and the guide RNA’s are inserted at separate genomic loci and depend on each other for spread. This is the safest way to develop and test gene drives in the laboratory to ensure no spread into the wild,” Akbari said.

Gene drives greatly increase the odds that a gene or set of genes will be passed on to offspring — from 50 percent to 99 percent. This number can potentially increase to 100 percent when a target gene is disrupted in multiple sites, a technique called multiplexing that has recently been mathematically modeled by Akbari and colleagues at UC Berkley.

Gene drives can be used to bias genetic inheritance in favor of rapidly spreading, self-destructive genes — such as those that disrupt fertility — and could be an environmentally friendly and cost-effective way to suppress populations of disease-spreading insects.

“Next steps should be undertaken to identify the regulatory sequences that can be used to express the guide RNAs from the genome, and once these sequences are identified developing gene drives in the species should be turnkey,” Akbari said.

Antibody protects against Zika and dengue, mouse study shows

Brazil and other areas hardest hit by the Zika virus – which can cause babies to be born with abnormally small heads – are also home to dengue virus, which is spread by the same mosquito species.

A new study led by researchers at Washington University School of Medicine in St. Louis shows that an antibody that protects against dengue virus is also effective against Zika in mice.

Antibodies remain in the bloodstream for weeks, so one or a few doses of an antibody-based drug given over the course of a woman’s pregnancy potentially could protect her fetus from Zika, with the added benefit of protecting her from both Zika and dengue disease, the researchers said. Dengue causes high fever, severe headaches, and joint and muscle pain in children and adults but does not directly harm fetuses.

“We found that this antibody not only neutralizes the dengue virus but, in mice, protects both adults and fetuses from Zika disease,” said Michael S. Diamond, MD, PhD, the Herbert S. Gasser Professor of Medicine and the study’s senior author.

The study is published Sept. 25 in Nature Immunology.

Since dengue and Zika are related viruses, the researchers reasoned that an antibody that prevents dengue disease may do the same for Zika. Diamond and graduate student Estefania Fernandez collaborated with Gavin Screaton, MD, DPhil, of Imperial College London, who had generated a panel of human anti-dengue antibodies years before.

The scientists infected nonpregnant adult mice with Zika virus and then administered one of the anti-dengue antibodies one, three or five days after infection. For comparison, another group of mice was infected with Zika virus and then given a placebo. Within three weeks of infection, more than 80 percent of the untreated mice had died, whereas all of the mice that received the anti-dengue antibody within three days of infection were still alive, and 40 percent of those that received the antibody five days after infection survived.

To find out whether the antibody also could protect fetuses from infection, the researchers infected female mice on the sixth day of their pregnancies with Zika virus and then administered a dose of antibody or a placebo one or three days later.

On the 13th day of gestation, the amount of Zika’s genetic material was 600,000 times lower in the placentas and 4,900 times lower in the fetal heads from the pregnant mice that were treated one day after infection, compared with mice that received the placebo. However, administering the antibody three days after infection was less effective: It reduced the amount of viral genetic material in the fetal heads nineteenfold and in the placentas twenty-threefold.

These findings suggest that for the antibody to effectively protect fetuses from Zika infection, it must be administered soon after infection. Such a goal may be unrealistic clinically because women rarely know when they get infected.

However, giving women the antibody as soon as they know they are pregnant could provide them with a ready-made defense against the virus should they encounter it. Antibody-based drugs have been used for decades to provide temporary protection against infectious diseases such as rabies when there is no time to vaccinate or, as in the case of Zika, when there is no vaccine available.

The key to using this antibody as a preventive drug would be to make sure that antibody levels in a woman’s bloodstream stay high enough to protect her fetus for the duration of her pregnancy.

Diamond and colleagues are working on identifying how much antibody a pregnant woman would need to ensure that her fetus is protected from Zika. They also are exploring ways to extend the antibody’s half-life in the blood, to reduce the number of times it would need to be administered.

Having anti-dengue antibodies circulating in the bloodstream for months on end poses a risk, though, because antibodies that protect against one strain of dengue virus sometimes worsen symptoms if a person is infected by another dengue strain.

To avoid the possibility of accidentally aggravating an already very painful disease, the researchers mutated the antibody in four spots, making it impossible for the antibody to exacerbate dengue disease.

“We mutated the antibody so that it could not cause antibody enhancement of dengue infection, and it was still protective,” said Diamond, who is also a professor of pathology and immunology, and of molecular microbiology. “So now we have a version of the antibody that would be therapeutic against both viruses and safe for use in a dengue-endemic area, because it is unable to worsen disease.”

AYUSH Medicines Developed for Mosquito-Borne Diseases like Dengue

The Research Councils viz Central Council for Research in Ayurvedic Science (CCRAS), Central Council for Research in Unani Medicine (CCRUM), Central Council for Research in Siddha (CCRS), Central Council for Research in Homeopathy (CCRH), autonomous bodies under the Ministry of AYUSH are engaged in Research and Development of new drugs. The details of new drugs developed by the Research Councils after clinical trials are as under: –

CCRAS:-

· AYUSH Rasayan A & B for geriatric health
· AYUSH A for Bronchial Asthma

· AYUSH-D for Diabetes Mellitus

· C1 oil for wound healing

· AYUSH-SL for Lymphatic Filariasis

· PJ-7 for dengue

· Carctol-S for Ovarian Cancer

· AYUSH M-3 for Migrain

CCRH:-

· Coleus forskohlii

· Catheranthus roseus

· Buxus sempervirens

· Cynara scolymus

· Hygrophilla spinosa

· Persea Americana

CCRS:-
· D5 chooranam for Diabetes mellitus,

· Rasagandhi mezhugu for Fibroid uterus,

· Sirupeelaiyathi Kudineer in Kalladaippu (Urolithiasis)

CCRUM:-

· Z. N. 5:- A herbal composition against bronchial asthma and process for preparation thereof.

· Cap. Habis:- A herbal composition effective against nazfuddam and a process for the preparation thereof.

· Cap. Mubarak:- A novel herbal composition effective as anti pyretic and to a process for the preparation thereof.

· Cap. Nazla:- A novel herbal composition effective against coryza and a process for preparing thereof.

· Qurs. Mafasil:- A novel herbal composition (sugar coated) effective against rheumatoid arthritis.

· Cap. Hudar:- A herbal composition effective against arthritis and to process for the preparation thereof.

· Qurs. Mulaiyin:- A novel herbal composition and a process for preparation thereof effective against constipation.

· Cap. Deedan:- A Novel Herbal Composition And A Process For Preparation Thereof Effective Against Abdominal Worm.

This information was given by the Minister of State (Independent Charge) for AYUSH, Shri ShripadYesso Naik in written reply to a question in Rajya Sabha today.